The “EZ4U assay cell viability assay” was highlighted in a recent study investigating the effects of RNA modifications (m6A demethylases) on renal cell carcinoma cell lines. Accumulating evidence shows that m6A RNA methylation participates in the pathogenesis of multiple diseases including cancers.
Renal cell carcinoma (RCC) accounts for approximately 3% of all cancers in adults. The 5-year survival rate for patients with locally advanced or metastatic RCC is poor with a prognosis of approximately 5%. Despite advanced pharmacological treatments of these patients (e.g. checkpoint immunotherapy, anti-VEGF antibodies, tyrosine kinase inhibitors), patient survival rates have only moderately improved.
Epigenetics studies the modification of heritable gene expression without having changes in the DNA sequence (1). N6-methyladenosine (m6A) is a common post-transcriptional methylation occurring in different types of RNAs in eukaryotic cells (2). m6A methylation is important in a number of physiological processes including the response to DNA damage. Recent studies suggest that RNA methylation has a great impact in cancer development and may provide a novel target for cancer treatment (3).
In a recent study researchers investigated the effects of RNA modifications (m6A demethylases) on renal cell carcinoma cell lines (4). Cell viability was determined with the BIOMEDICA EZ4U (easy for you) assay. In brief, cells were incubated with the EZ4U substrate for 3 hours at 37°C and absorbance was measured at 450 nm with 620 nm as reference.
Measuring Cell Viability with the EZ4U (MTT) Assay (developed and manufactured by Biomedica)
The EZ4U (easy for you) assay is a metabolic assay (similar to the MTT assay) that can quantify cell health. It measures the reduction of the colorimetric substrate through the activity of mitochondrial enzymes in the cells.
Assays measuring cell viability are a widely used tool in cell biology research. The EZ4U test kit is a straightforward and reliable non-radioactive assay that is completed within two to five hours, depending on the cell type studied. The EZ4U assay employs non-toxic tetrazolium salts, which are, during incubation, converted into colored formazan. As the reduction process relies on functional mitochondria that become inactive shortly after cell death, this method effectively distinguishes between living and dead cells. The assay procedure is identical to the thymidine incorporation procedure. Therefore, no changes in test protocols are necessary. Furthermore, an additional benefit is the ability to continue cultivation after determining cell numbers.
The EZ4U cell proliferation and cytotoxicity assay is highly suited for testing cell viability in a number of different cell types. The assay has been used in various cell types and citations can be found in the link.
EZ4U Cell Proliferation and Cytotoxicity Assay –single incubation step using living cells
Brief description: 20µl of the EZ4U substance (colorless non-toxic tetrazolium salts) is first added to each well. The mitochondria of the living cells oxidize the EZ4U substance to a yellow-colored formazan derivate. The color change is measured on a conventional plate reader at 450 nm using a reference wavelength of 620 nm.
EZ4U Cell Viability & Cytotoxicity Assay (cat.no. BI-5000)
- Reliable & Sensitive
- One step incubation – for use on living cells
- Widely used – referenced in +270 publications
Download the BROCHURE – EZ4U cell proliferation and cytotoxicity assay and the PROTOCOL BOOKLET
Literature
- The role of m6A modification in the biological functions and diseases. Jiang X et al., Signal Transduct Target Ther. 2021; 21;6(1):74. PMID: 33611339.
- Depletion of the m6A demethylases FTO and ALKBH5 impairs growth and metastatic capacity through EMT phenotype change in clear cell renal cell carcinoma. Hu W et al., Am J Transl Res. 2023; 15(3):1744-1755. PMID: 37056835.
- The potential role of RNA N6-methyladenosine in Cancer progression. Wang T et al., Mol Cancer. 2020. 19(1):88. PMID: 32398132.
- Depletion of the m6A demethylases FTO and ALKBH5 impairs growth and metastatic capacity through EMT phenotype change in clear cell renal cell carcinoma. Hu W Am J Transl Res. 2023 Mar 15;15(3):1744-1755. PMID: 37056835.
ELISA (Enzyme Linked Immunosorbent Assay) tests are one of the most used biomedical methods for quantifying analytes in biological samples. Apart from their sensitivity and specificity, they are characterized by their simplicity and cost-effectiveness compared to many other analytical methods. However, the easiness in performing an ELISA assay may mask some of the more complex aspects when measuring biological samples.
One of the most common ELISA techniques is the Sandwich ELISA which is used to measure the amount of antigen between two layers of antibodies. Sandwich ELISAs have the limitation that the antigen (e.g. protein in the sample) must have at least two antigen binding sites so that at least two different antibodies can bind. However, these assays are useful when the concentration of the antigen in the sample is low or high concentrations of other antigens contaminate a sample.
Principle of a Sandwich ELISA Assay
Typically a 96-well microtiter plate is pre-coated with a specific “capture antibody” (monoclonal or polyclonal antibody). After adding the sample to the microtiter plate and during an incubation period, the protein of interest present in the sample binds to the pre-coated antibody in the well. Next, the “detection antibody” is added to the well, which forms a sandwich (capture antibody – target antigen – detection antibody).
Figure 1: ELISA Assay Principle
The results can be quantified by adding an enzyme conjugated antibody specific for the second epitope, catalyzing an enzymatic color reaction. The amount of bound enzyme conjugate in the well is detected after following a short incubation with an appropriate substrate. The resulting signal (color change in the well) is detectable with a standard microtiter plate ELISA reader measuring the absorbance (optical density) – the greater the signal, the greater the concentration of the analyte in the sample that is measured throughout the range of the standard curve.
Figure 2: Typical standard curve (using 7 calibrators) for a sandwich ELISA assay, showing the relation between the signal and the analyte concentration
How to Obtain Reproducible ELISA Assay Results
To obtain reliable results it is recommended to choose an ELISA Assay that has gone through a full validation protocol. Results (validation data) must be provided that include data on specificity, sensitivity (LOD, LLOQ), accuracy/recovery, parallelism and dilution linearity, precision (within-run and in-between run), calibration, stability, and lot-to-lot consistency.
CORNERSTONES OF AN ELISA ASSAY VALIDATION
1.SPECIFICITY
Ensuring accurate measurement of the protein of interest
The performance of an ELISA is linked to the quality of the antibody pairs used for the analyte/biomarker detection.
The supplier (assay developer ) needs to :
-select antibody pairs with high affinity and specificity with mapped binding sites
-optimize the ELISA kits to quantify biomarkers in both healthy and pathological samples
2.SENSITIVITY
Enabling the detection of low levels of the analyte of interest
The sensitivity of an ELISA assay refers to the lowest limit of detection (LOD) of the protein that can be detected with the antibody pair used in the ELISA kit. The sensitivity depends mainly on the affinity of the solid phase antibody (coating antibody). Therefore, using a high affinity antibody can increase sensitivity.
-Analytical sensitivity – limit of detection (LOD) is the lowest concentration that can be measured (detected) with statistical significance by means of a given analytical procedure. This concentration is calculated as the background +/- 2 standard deviations.
-Functional Sensitivity – lower limit of detection (LLOQ) is the lowest concentration at which the analyte can be reliably detected.
3.ACCURACY / RECOVERY
Ensuring the correct presence or absence of the protein of interest in samples with different matrices (serum, plasma)
The accuracy in an ELISA assay correctly identifies the presence or absence of the target protein / antigen (biomarker of interest) in a specific sample, which excludes matrix effects that may interfere with the measurement of the analyte of interest.
-Ideally, the accuracy should be determined in all of the sample types (e.g. serum, EDTA plasma, heparin plasma..) where the analyte of interest will be measured. For this reason, the various sample matrices are spiked with known amounts of the recombinant analyte. Samples are analysed against the standard/calibration curve of the assay and then compared with the nominal value.
4.PARALLELISM / DILUTION LINEARITY
Ensuring that samples can be measured with confidence / Ensuring that serial dilutions of samples are quantitated accurately
-Parallelism determines whether real samples with high endogenous analyte concentrations yield the same level of detection in the standard curve after sample dilution. This procedure highlights differences in the binding affinity of the antibodies to the endogenous analyte and the analyte used for calibrating the ELISA. Parallelism in real samples containing endogenous levels of the analyte should always be tested during assay validation.
-Dilution linearity determines the recovery of the analyte in samples that are spiked with the recombinant analyte, used for calibrating the assay (standards).
5.PRECISION
Ensuring precise and consistent results within and across lots
-Within-run precision (repeatability) – evaluation of the precision of the ELISA that measures the variation between multiple determinations of one sample in one single test run.
-In-between run precision – evaluation of samples that are tested several times within one ELISA assay lot guaranteeing accurate results when using ELISA kits that derive from different kit lots.
6.CALIBRATION
Enabling the accurate quantification of the protein of interest
-The accurate quantification depends on the linearity and the reproducibility of the standard curve. During the optimization process of the ELISA kit, low variability between the results of the calibrators must be ensured. Whenever available, standard calibration with standards calibrated to NIBSC standards or WHO reference should be employed to ensure a harmonized standardization.
7.STABILITY
Enabling stability of the analyte of interest in respective samples as well as the stability of the reagents in the ELISA kit
During development, stability of all assay components as well as the stability of the analyte of interest in the respective sample matrices (serum, plasma) should be tested.
-The stability of the analyte can be tested in samples when e.g. samples undergo repeated freeze-thaw cycles or if samples are exposed to room-temperature for a prolonged (defined) period of time.
-Assay kit stabilty (e.g. stability of the kit reagents) should be tested at various temperature settings for a defined period of time.
-Shelf-life of the kit: the real-time stabilty of kit reagents should be tested during the shelf life of the kit. e.g. when kit reagents are stored at 4°C for 6/12/18 months.
8.LOT-TO-LOT CONSISTENCY
Ensuring that consistent results are obtained with different ELISA kit lots
The use of “in-house controls” (serum and plasma samples) containing the endogenous analyte as well as samples that are spiked with the recombinant analyte are tested in every kit lot batch to evaluate whether the data are within the set quality control ranges.
How to Obtain Reproducible ELISA Assay Results
- BIOMEDICA´s ELISA kits undergo a full validation following international quality guidelines making sure that your precious samples only detect the analyte of interest.
Learn more about how to obtain reproducible ELISA assay results by clicking on the link:
WHY CHOOSE ELISA KITS from BIOMEDICA?
Specific – accurate biomarker detection with characterized antibodies
Reliable –validation using clinical samples
Reproducible – guaranteed lot to lot consistency
Further reading
A Practical Guide to Immunoassay Method Validation. Andreasson Uet al., Front Neurol. 2015 Aug 19;6:179. doi: 10.3389/fneur.2015.00179. PMID: 26347708; PMCID: PMC4541289.
IL-6 (Interleukin-6) is a protein that acts both as a pro-inflammatory cytokine and as an anti-inflammatory myokine (1). During physical activity, IL-6 is released from muscle fibers into the bloodstream where it has different functions:
–Energy Balance: IL-6 is a regulator of energy metabolism. It promotes the breakdown of fats and stimulates glucose production, thus ensuring that muscles have a constant energy supply during prolonged exercise.
–Anti-inflammatory effects: Although IL-6 is an immunoregulatory molecule that promotes inflammation in certain contexts, IL-6 also has anti-inflammatory properties during exercise. It stimulates the production of anti-inflammatory cytokines and inhibits the production of pro-inflammatory cytokines, reducing overall inflammation in the body.
–Muscle repair: IL-6 activates pathways that contribute to muscle regeneration and adaptation, thus helping muscles to recover and become stronger after physical activity.
Interleukin-6 (IL-6) and Exercise
The efficacy of exercise to reverse frailty in an aging population has been investigated in a randomized controlled trial (2). The study investigated the effectiveness of a multicomponent exercise program (MCEP) in older adults that also included the measurement of blood biomarkers Interleukin-6 (IL-6) and C-reactive protein (CRP). The results clearly indicate that when compared with controls at 12-weeks, the MCEP group significantly showed a decrease in IL-6 and CRP levels (p < 0.05). In conclusion, the study clearly demonstrated that the MCEP exercise program was effective in improving physical performance and quality of life. Further, it delays frailty and decreases inflammation in frail older adults (2).
IL-6 can easily be measured in various biological fluids with a conventional ELISA assay.
Human IL-6 ELISA (cat. no. BI-IL6)
- HIGH SENSITIVITY – detectable values in serum and plasma samples
- RELIABLE – full validation
- CONVENIENT – ready to use calibrators and controls
- PROMOTION – Take 15% off!
IL-6 ELISA ASSAY CHARACTERISTICS (#BI-IL6)
- Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
- Sample type: Plasma (EDTA, citrate, heparin), serum, cell culture, and urine
- Sample volume / well: 100 µl
- Incubation: 2 h / 1 h / 1 h / 30 min
- Sensitivity: 0.28 pg/ml (LLOQ: 0.78 pg/ml (measurable concentrations in serum AND plasma samples)
- Assay range: 0 – 200 pg/ml (ready standards with concentrations at: 0 /3.12 / 6.25 / 12.5 / 25 / 25 / 50 /100 /200 pg/ml)
- Precision: In-between-run (n=3): ≤ 6 % CV, within-run (n=3): ≤ 7 % CV
- Specificity: The human IL-6 ELISA recognizes recombinant and endogenous (natural human IL-6)
- Validation data: precision, accuracy, dilution linearity, parallelism, values for healthy donors are shown here
- Use: research use only
Literature
- Interleukin-6 myokine signaling in skeletal muscle: a double-edged sword? Muñoz-Cánoves Pet al., FEBS J. 2013; 280(17):4131-48. doi: 10.1111/febs.12338.
- Multicomponent Exercise Program Reduces Frailty and Inflammatory Biomarkers and Improves Physical Performance in Community-Dwelling Older Adults: A Randomized Controlled Trial. Sadjapong U et al., Int J Environ Res Public Health. 2020; 7(11):3760. doi: 10.3390/ijerph17113760.
We are present in over 60 countries, ensuring global availability and customer support through partnerships with selected local distributors on almost every continent. Regardless of your location, we are here to help you reach your goals.
With over 30 years of experience, we specialize in developing and manufacturing high-quality ELISA assay kits for biomarkers in clinical research.
Check out our ELISA Assay product brochure
Our ELISA Kits Travel Worldwide – contact us or your local representative
About us
BIOMEDICA develops and manufactures high quality and widely cited immunoassays for clinical and pre-clinical applications in bone and cardiorenal diseases. For specific biomarkers, Biomedica has become a world-wide leader, e.g. Sclerostin, free sRANKL, OPG, DKK-1, and NT-proCNP.
All Biomedica ELISA assays are fully validated following international quality guidelines. The assays include ready-to-use serum-based calibrators and controls enabling researchers to collect biologically reliable data.
This year’s ELISA assay highlights
Cardiac Safety Biomarkers : Rat NT-proBNP, NT-proANP ELISA kits
FGF23 : FGF23 intact , FGF23 (C-terminal) ELISA kits
Cytokines : VEGF, Angiopoietin-2, IL-6
Discover our PROMOTION : Take 15% OFF of the human IL-6 ELISA Assay now through end of 2024! Making IL-6 ELISA Kits more affordable!
Features & Benefits of BIOMEDICA ELISA Assay Kits
- Highly Specific – many of our kits employ characterized epitope-mapped antibodies
- Excellent Quality – our kits are validated according to international quality guidelines (ICH, FDA..)
- Convenient Use –offering ready to use and color coded reagents – controls are included in all kits!
Other Products
EZ4U – Cell Proliferation & Cytotoxicity Assay (cat. no. BI-5000) for reliable testing of cell viability in 96-well microtiter plates.
Features & Benefits EZ4U Assay
-easy and convenient singe-step incubation for use on living cells
-non-radioactive
-cultivation can be continued after cell numbers are determined
-widely cited in more than 240 publications e.g.
- BRD4-targeting PROTACs Synergize With Chemotherapeutics Against Osteosarcoma Cell Lines
Lang et al., Anticancer Res; 2024; 44 (3), 971-980. PMID: 38423674
- Expression of cytokines in pleural effusions and corresponding cell lines of small cell lung cancer
Rath et al., Transl Lung Cancer Res; 2024; 13 (1), 5-15. PMID: 38405004
IL-6 is a pleiotropic cytokine that is involved both in immune response and in inflammation, hematopoiesis, bone metabolism, and embryonic development (1). IL-6 plays roles in chronic inflammation that are closely related to chronic inflammatory diseases and autoimmune diseases (2).
Paradoxical effects of IL-6 in preventing or promoting cancer development
A recent review highlights the paradoxical effects of IL-6 in preventing or promoting cancer development (3). During exercise, IL-6 is released from working muscle fibers, enabling the communication between muscles and other organs to maintain energy balance and providing anti-inflammatory benefits (4). It has been shown that muscle-derived IL-6 improves insulin sensitivity in glycogen-storing tissues, hereby stimulating the release of anti-inflammatory cytokines in the blood, mobilizing cytotoxic immune cells, and decreasing DNA damage in cancer cells. These effects of IL-6 may help protect against cancer development and progression.
On the other hand, IL-6 is also induces inflammatory effects by activating the transcription of factors across various inflammatory pathways. It is secreted by immune cells at sites of inflammation and within the tumor environment. Continuous IL-6 signaling at sites of inflammation and within the tumor microenvironment leads to chronic low-grade inflammation and activates pathways that support tumor growth (2, 5).
Thus, although IL-6 mostly promotes tumor growth and progression, it can also exhibit dual effects depending on the context, that is particularly related to the tumor environment and stage. It can stimulate an anti-tumor immune response by activating immune cells, supporting immune surveillance and targeting pre-cancerous cells.
Features & Benefits of BIOMEDICA´s human IL-6 ELISA Assay (#BI-IL6)
- High sensitivity – measurable concentrations in serum AND plasma samples
- Reliable- rigorously validated following international quality guidelines
- Simple – ready to use color coded reagents (7 pre-diluted standards, 2 controls)
- Standardized – calibration using WHO standard
Human Interleukin–6 (IL-6) ELISA ASSAY CHARACTERISTICS
- Catalog Number: BI-IL6
- Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
- Sample type: Serum, plasma (EDTA, citrate, heparin), cell culture supernatatants, urine
- Sample volume: 100 µl / well
- Assay time: 2 h / / 1 h / 1 h / 30 min
- Sensitivity: LOD 0.28 pg/ml; LLOQ: 0.78 pg/ml (measurable concentrations in serum AND plasma samples)
- Standard range: 0 – 200 pg/ml (0 /3.12 / 6.25 / 12.5 / 25 / 25 / 50 /100 /200 pg/ml)
- Precision: In-between-run (n=3): ≤ 6 % CV, within-run (n=3): ≤ 7 % CV
- Specificity: The human IL-6 ELISA recognizes recombinant and endogenous (natural human IL-6)
- Validation data: precision, accuracy, dilution linearity, parallelism, values for healthy donors are shown here
- Use: research use only
Click here for more information on the human IL-6 ELISA
Literature
- Historical overview of the interleukin-6 family cytokine. Kang S, Narazaki M, Metwally H, Kishimoto T J Exp Med. 2020 May 4;217(5):e20190347. doi: 10.1084/jem.20190347. Erratum in: J Exp Med. 2020 May 4;217(5):jem.2019034704212020c. doi: 10.1084/jem.2019034704212020c. PMID: 32267936; PMCID: PMC7201933.
- IL-6 in inflammation, autoimmunity and cancer. Hirano T. Int Immunol. 2021 Mar 1;33(3):127-148. doi: 10.1093/intimm/dxaa078. PMID: 33337480; PMCID: PMC7799025.
- The exercise IL-6 enigma in cancer. Orange ST, Leslie J, Ross M, Mann DA, Wackerhage H.Trends Endocrinol Metab. 2023 Nov;34(11):749-763. doi: 10.1016/j.tem.2023.08.001. Epub 2023 Aug 24. PMID: 37633799.
- IL-6 signaling in acute exercise and chronic training: Potential consequences for health and athletic performance. Nash D, Hughes MG, Butcher L, Aicheler R, Smith P, Cullen T, Webb R. Scand J Med Sci Sports. 2023 Jan;33(1):4-19. doi: 10.1111/sms.14241. Epub 2022 Oct 8. PMID: 36168944; PMCID: PMC10092579.
- Interleukin-6 in Hepatocellular Carcinoma: A Dualistic Point of View. Nenu I, Toadere TM, Topor I, Țichindeleanu A, Bondor DA, Trella ȘE, Sparchez Z, Filip GA. Biomedicines. 2023 Sep 24;11(10):2623. doi: 10.3390/biomedicines11102623. PMID: 37892997; PMCID: PMC10603956.
July is acknowledged as Bone Cancer and Sarcoma Awareness Month, focusing on increasing awareness about rare and challenging cancers affecting children and adolescents. Sarcoma is a type of cancer that originates in the bones and soft tissues. Soft tissues include muscle, fat, blood vessels, fibrous tissues such as tendons and ligament. More than 70 different subtypes of sarcomas have been reported. However, all sarcomas can be divided into two main groups: soft tissue sarcomas and bone cancers.
Soft Tissue Sarcomas is a rare type of cancer that originates from the growth of cells within the body´s soft tissue. More than 50 different soft tissue sarcomas have been reported (3 ) Soft tissue sarcoma can occur anywhere in the body, but it most frequently develops in arms, legs, and abdomen (4).
Osteosarcoma is the most prevalent type of bone cancer in children and adolescents with and incidence of around 4.4. cases per million children reported each year (1). Genomic alterations, particularly the inactivation of TP53 and RB, are present in most cases of osteosarcoma.
Ewing sarcoma is the second most common bone tumor occurring most frequently in teenagers, with a median age of 15 years. Ewing sarcomas is an aggressive tumor that develops usually in bone, but sometimes in soft tissue, most commonly affecting the lower extremity and pelvis. At diagnosis, up to 25% of patients , commonly found in the lung, bones, and bone marrow (2). This condition is biologically driven by a chromosomal translocation, typically involving the EWS and FLI1 genes.
July is Sarcoma and Bone Cancer Awareness Month
- 5 Things to Know About Bone Cancer and Sarcoma Awareness Month- link
Wnt Signalling molecules Sclerostin (SOST) and Dickkopf-1 (DKK-1) in Sarcoma
SCLEROSTIN has been shown to be expressed in bone and cartilage forming skeletal tumors. Sclerostin is widely localized to areas in osteoblastic differentiation and ossification (5). In a study using an osteosarcoma mouse model, the administration of sclerostin resulted in inhibited tumor growth and extended survival periods (6).
DKK-1 is a wnt inhibitor that has been shown to partially improve osteosarcoma survival by upregulating aldehyde-dehydrogenase-1A1, neutralizing reactive oxygen species originating from nutritional stress and chemotherapeutic challenge (7). In a mouse model researches demonstrated the use of a DKK-1 targeting vivo morpholino that reduces tumour progression (7).
FGF23 in uterine sarcoma
Human FGF23 has been shown to be highly expressed in uterine sarcoma highlighting its potential as a biomarker for the diagnosis and prognosis of the disease (8).
Sclerostin, DKK-1 and FGF23 can easily be measured in blood, urine and cell culture supernatants with an ELISA assay.
– Sclerostin ELISA (cat. no. BI-20492)
– DKK-1 ELISA (cat. no. BI-20413)
– FGF23 ELISAs (cat. no. BI-20700, BI-20702)
Literature
- Osteosarcoma. Pediatr Blood Cancer. Eaton BR, Schwarz R, Vatner R, Yeh B, Claude L, Indelicato DJ, Laack N. 2021 May;68 Suppl 2:e28352. doi: 10.1002/pbc.28352. Epub 2020 Aug 11. PMID: 32779875.
- Ewing sarcoma. Pediatr Blood Cancer. Eaton BR, Claude L, Indelicato DJ, Vatner R, Yeh B, Schwarz R, Laack N. 2021 May;68 Suppl 2:e28355. doi: 10.1002/pbc.28355. PMID: 33818887.
- More Than 50 Subtypes of Soft Tissue Sarcoma: Paving the Path for Histology-Driven Treatments. Katz D, Palmerini E, Pollack SM. Am Soc Clin Oncol Educ Book. 2018 May 23;38:925-938. doi: 10.1200/EDBK_205423. PMID: 30231352.
- Malignant Soft-Tissue Sarcomas. Hematol Oncol Clin North Am. Brownstein JM, DeLaney TF. 2020 Feb;34(1):161-175. doi: 10.1016/j.hoc.2019.08.022. Epub 2019 Oct 28. PMID: 31739942.
- Sclerostin expression in skeletal sarcomas. Shen J, Meyers CA, Shrestha S, Singh A, LaChaud G, Nguyen V, Asatrian G, Federman N, Bernthal N, Eilber FC, Dry SM, Ting K, Soo C, James AW. Hum Pathol. 2016 Dec;58:24-34. doi: 10.1016/j.humpath.2016.07.016. Epub 2016 Aug 3. PMID: 27498059; PMCID: PMC6560186.
- Antitumor Effect of Sclerostin against Osteosarcoma. Ideta H, Yoshida K, Okamoto M, Sasaki J, Kito M, Aoki K, Yoshimura Y, Suzuki S, Tanaka A, Takazawa A, Haniu H, Uemura T, Takizawa T, Sobajima A, Kamanaka T, Takahashi J, Kato H, Saito N. Cancers (Basel). 2021 Nov 29;13(23):6015. doi: 10.3390/cancers13236015. PMID: 34885123; PMCID: PMC8656567.
- Morpholino-driven blockade of Dkk-1 in osteosarcoma inhibits bone damage and tumour expansion by multiple mechanisms. Pan S, Cesarek M, Godoy C, Co CM, Schindler C, Padilla K, Haskell A, Barreda H, Story C, Poole R, Dabney A, Gregory CA. Br J Cancer. 2022 Jul;127(1):43-55. doi: 10.1038/s41416-022-01764-z. Epub 2022 Mar 11. PMID: 35277659; PMCID: PMC9276700.
- Fibroblast Growth Factor 23 is a Potential Prognostic Biomarker in Uterine Sarcoma. Yang L, Cai Y, Wang Y, Huang Y, Zhang C, Ma H, Zhou JG. Technol Cancer Res Treat. 2024 Jan-Dec;23:15330338241245924. doi: 10.1177/15330338241245924. PMID: 38613349; PMCID: PMC11015760.
We´re excited to offer our customers a significant price drop by 15% on our
human Interleukin-6 ELISA kit (cat. no. BI-IL6)
Quality and fully validated assay with ready to use reagents for affordable and efficient research!
⇒ Contact us or our local representative
Making IL-6 ELISA Kits More Affordable
Take 15% OFF of the human IL-6 ELISA Assay now through end of 2024!
BIOMEDICA´s human Interleukin-6 (IL-6) ELISA Assay is highly sensitive and shows detectable values in serum and in plasma samples.
Features & Benefits
- High Specificity – using characterized epitope-mapped antibodies
- High Sensitivity – detectable values in both serum and plasma
- Superior Quality – validated according to international quality guidelines
- Standardized – use of WHO intern.standards for kit calibration & harmonization
- User-Friendly –ready to use color coded prediluted standards & controls
BIOMEDICA Human IL-6 ELISA kit
- Product code: BI-IL6
- Method: ELISA
- Time to result: 4.5 hours
- Sample types: human serum, plasma (EDTA, citrate, heparin), cell culture supernatants, urine
- Sample volume: 100 µl/well
- Sensitivity LOD: 0.28 pg/ml (measurable concentrations in serum AND plasma samples!)
- Standard curve range: 0 – 200 pg/ml (0 / 3.12 / 6.25 / 12.5 / 25/ 50 / 100 / 200)
- Specificity: Endogenous and recombinant human IL-6
- Protokol booklet
- Validation data file
About Interleukin-6
Interleukin-6 (IL-6) is a multifunctional cytokine that plays an important role in the body´s response to tissue injury or infection. Il-6 is produced by various cells, including T-cells, B-cells, macrophages, as well as vascular endothelial cells, mast cells, and dentritic cells (1). Il-6 secretion is stimulated during inflammatory response and travels through the bloodstream to the liver, where it induces the production of acute phase reactants such as C-reactive protein and others (1).
Clinical significance of Interleukin-6
-Inflammatory diseases: elevated IL-6 levels are detected in various inflammatory conditions such as rheumatoid arthritis (2), systemic lupus erythematosus (3) , and inflammatory bowel disease (4). Therapies targeting Il-6 have been already approved (5).
-Cancer: Il-6 is abundantly present in the tumor microenvironment of various tumours. It promotes tumnorigenesis, invasiveness, and metastases in cancer (6). New perspectives in cancer immunotherapy targeting IL-6 are discussed (7).
-Metabolic diseases: Dysregulated IL-6 activity is associated with pathologies involving chronic inflammation and autoimmunity including metabolic diseases like obesity, metabolic syndrome or type-2 diabetes (8).
Literature
- Interleukin 6: at the interface of human health and disease. Grebenciucova E et al., Front Immunol. 2023; 28;14:1255533. PMID: 37841263.
- Interleukin-6 in Rheumatoid Arthritis. Pandolfi F et al., Int J Mol Sci. 2020 ; 23;21(15):5238. PMID: 32718086; PMCID:.
- Role of IL-6 and IL-6 targeted therapy in systemic lupus erythematosus. Nepal D, Gazeley D. Rheumatology (Oxford). 2023; 1;62(12):3804-3810. PMID: 37594751.
- Cytokines of the interleukin-6 family as emerging targets in inflammatory bowel disease. Garbers C, Lokau J. Expert Opin Ther Targets. 2024; 28(1-2):57-65. PMID: 38217849.
- Therapeutic uses of anti-interleukin-6 receptor antibody. Kang S et al., Int Immunol. 2015; 27(1):21-9. PMID: 25142313.
- The Role of IL-6 in Cancer Cell Invasiveness and Metastasis-Overview and Therapeutic Opportunities. Rašková M et al., Cells. 2022; 21;11(22):3698. PMID: 36429126.
- New perspectives in cancer immunotherapy: targeting IL-6 cytokine family. Soler MF et al., J Immunother Cancer. 2023; 11(11):e007530. PMID: 37945321.
- IL-6 family cytokines as potential therapeutic strategies to treat metabolic diseases. Zhao J et al., Cytokine. 2021; 144:155549. PMID: 33962843
Fibroblast growth factor 23 (FGF23) is a hormone primarily known for its role in mineral metabolism, particularly in regulating phosphate homeostasis.
FGF23 is primarily produced in the bone by osteocytes and acts on the kidneys to regulate phosphate reabsorption and vitamin D metabolism. FGF23 helps to maintain phosphate levels within the normal range by promoting urinary phosphate excretion and inhibiting the production of vitamin D (1). The dysregulation of FGF23 can lead to conditions such as hypophosphatemic rickets and hyperphosphatemia.
FGF23 and its effects on the heart
Numerous clinical studies have shown that elevated plasma level concentrations of FGF23 are linked to left ventricular hypertrophy (LVH), heart failure, and increased mortality in the general population, particularly among individuals with chronic kidney disease (CKD) (2). The exact mechanisms by which FGF23 affects the heart are still under investigation, but several possible pathways have been suggested. For instance, FGF23 may induce hypertrophy directly through binding to specific FGF23 receptors expressed by cardiomyocytes in the heart. FGF23 may also promote hypertrophy indirectly by altering mineral metabolism, leading to increased circulating levels of phosphorus, which can contribute to cardiovascular damage.
In addition, FGF23 may have effects on the endothelium, the inner lining of blood vessels. It has been suggested that FGF23 may hinder endothelial function and promote vascular calcification, thereby contributing to the development of cardiovascular disease. Further research is needed to fully understand the exact mechanisms through which FGF23 affects the heart and its role in cardiovascular disease (3).
Measurement of FGF23
FGF23 concentrations can be measured with blood tests. The most common way for measuring FGF23 is with a conventional ELISA assay (enzyme-linked immunosorbent assay). The ELISA utilizes specific antibodies that recognize and bind FGF23 molecules present in the sample. The intensity of the antibody-antigen reaction is measured, allowing an easy quantification of FGF23 in the respective sample.
Currently there are two different assays that allow the measurement of FGF23 in blood samples:
Intact FGF23 Assays
intact FGF23 represents the full length, biologically active form of the hormone. It consists of the complete FGF23 protein structure without being enzymatically cleaved.
The BIOMEDICA FGF23 (intact) human ELISA (cat. no. BI-20700)
is a sandwich-based immunoassay with an anti-human FGF23 capture antibody recognizing a structural epitope in the N-terminal part of FGF23 and a detection antibody binding to the C-terminal part of mature FGF23.
FGF23 C-terminal Assays
The c-terminal fragments of FGF23 are the result of the enzymatic cleavage of the intact FGF23 molecule.
Of note: all commercially available FGF23 c-terminal assays detect both c-terminal FGF23 fragments as well as the intact FGF23 molecule!
The BIOMEDICA FGF23 (C-terminal) human multi-matrix ELISA (cat. no. BI-20702)
is a sandwich-based immunoassayw hich recognizes multiple epitopes in the C-terminal part of FGF23.
Features & Benefits
- Validated ELISA kits following international quality guidelines
- Biomedica´s FGF23 assay have been used in numerous studies
validation reports and citations can be downloaded here
Literature
- Biology of Fibroblast Growth Factor 23: From Physiology to Pathology. Courbebaisse M, Lanske B. Cold Spring Harb Perspect Med. 2018 May 1;8(5):a031260. doi: 10.1101/cshperspect.a031260. PMID: 28778965; PMCID: PMC5932574.
- Direct and indirect effects of fibroblast growth factor 23 on the heart. Nakano T, Kishimoto H, Tokumoto M Front Endocrinol (Lausanne). 2023 Feb 24;14:1059179. doi: 10.3389/fendo.2023.1059179. PMID: 36909314; PMCID: PMC9999118.
- FGF23 directly impairs endothelium-dependent vasorelaxation by increasing superoxide levels and reducing nitric oxide bioavailability. Silswal N, Touchberry CD, Daniel DR, McCarthy DL, Zhang S, Andresen J, Stubbs JR, Wacker MJ. Am J Physiol Endocrinol Metab. 2014 Sep 1;307(5):E426-36. doi: 10.1152/ajpendo.00264.2014. Epub 2014 Jul 22. PMID: 25053401; PMCID: PMC4154070.
for ELISA and Luminex Assays and microRNA Analysis
Biomedica Immunoassays is dedicated to developing and producing high-quality ELISA assay kits.
Building on this foundation, we also specialize in providing customized sample testing services for customers in academia, biotechnology, and the pharmaceutical industries.
Sample Testing Services
for ELISA and Luminex Assays and microRNA Analysis
Share the details of your project with us! At Biomedica, our scientists will consult with you to discuss your project and provide scientific and/or technical support for your ongoing research, ensuring reliable and relevant results. We will supply you with raw data as well as a professionally formatted analytical report.
We offer:
- Customized Services – Flexibility to meet your project needs
- Expertise – Trained and experienced laboratory staff
- Quality Assurance – High-quality equipment adhering to strict quality guidelines
- Timeliness – Rapid turn-around times to meet your deadlines
- Reliability – Verified results with comprehensive analytical reports
Sample Testing Services for ELISA and Luminex Assays and microRNA Analysis
We will measure your clinical or pre-clinical samples for any selected biomarker using ELISA (enzyme-linked immunosorbent assay) or Luminex.
- Clinical Samples: serum, plasma, urine, cell culture supernatants
- Pre-Clinical Samples: mouse, rat, rabbit, or other species (e.g., NT-proBNP or NT-proANP measurement)
- Custom Applications: contact us for tailored solutions
Click here to learn about the workflow. Our measurement services include:
ELISA Assay Kits
We offer services for our proprietary Biomedica assays (see product list) as well as ELISA Assay Kits from other providers, covering a wide range of biomarker analytes.
Luminex Technology Multiplex Assays
We utilize Luminex xMAP® (multiple analyte profiling) technology-based immunoassays to measure your samples, enabling the simultaneous detection and quantification of multiple biomarkers.
For additional details, please refer to our workflow chart or reach out to us directly to discuss how we can support your specific research project
NEXT-GENERATION SEQUENCING & RT-qPCR MICRORNA SERVICES
We provide a comprehensive range of high-quality RNA services, including RNA extraction, next-generation sequencing (NGS), RT-qPCR, and custom analysis of microRNA signatures, all performed by our experienced laboratory staff. Our services include:
- RNA extraction from biofluids (serum, plasma, extracellular vesicles), cells, and tissues (quality control of total RNA utilizes Bioanalyzer chips).
- Next-generation sequencing (NGS).
- RT-qPCR.
- Cell-type specific microRNA/mRNA analysis in complex tissues, along with custom analysis of microRNA signatures.
For more details about our microRNA services, please visit our website or contact us directly.
Further reading
Enzyme-Linked Immunosorbent Assay: Types and Applications. Hayrapetyan H et al., Methods Mol Biol. 2023;2612:1-17. doi: 10.1007/978-1-0716-2903-1_1. PMID: 36795355.
A comprehensive review of Dynamic Chemical Labelling on Luminex xMAP technology: a journey towards Drug-Induced Liver Injury testing. Marín-Romero A, Pernagallo S Anal Methods. 2023 Nov 23;15(45):6139-6149. doi: 10.1039/d3ay01481a. PMID: 37965948.
Circulating microRNAs as novel biomarkers for bone diseases – Complex signatures for multifactorial diseases? Hackl M et al., Mol Cell Endocrinol. 2016 Sep 5;432:83-95. doi: 10.1016/j.mce.2015.10.015. Epub 2015 Oct 23. PMID: 26525415.
For affordable and efficient research
Interleukin-6 (IL-6) is a multifunctional cytokine that plays an important role in the body´s response to tissue injury or infection. Il-6 is produced by various cells, including T-cells, B-cells, macrophages, as well as vascular endothelial cells, mast cells, and dentritic cells (1). Il-6 secretion is stimulated during inflammatory response and travels through the bloodstream to the liver, where it induces the production of acute phase reactants such as C-reactive protein and others (1).
Human Interleukin-6 ELISA Assay Kit – high sensitivity
BIOMEDICA´s human Interleukin-6 (IL-6) ELISA Assay is highly sensitive and shows detectable values in serum and in plasma samples.
Features & Benefits
- High Specificity – using characterized epitope-mapped antibodies
- High Sensitivity – measurable values in both serum and plasma
- Superior Quality – validated according to international quality guidelines
- Standardized – use of WHO intern.standards for kit calibration & harmonization
- User-Friendly – ready to use color coded prediluted standards & controls
Take 15% off the human IL-6 ELISA Assay now through end of 2024!
Use Promotion Code: BCIL62024 on your order.
Human IL-6 ELISA Kit (cat. no. BI-IL6)
- Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
- Sensitivity LOD: 0.28 pg/ml (measurable concentrations in serum AND plasma samples!)
- Sample types: human serum, plasma (EDTA, citrate, heparin), cell culture supernatants, urine)
- Sample volume: 100 µl/well
- Dynamic range: 0 – 200 pg/ml (0 / 3.12 / 6.25 / 12.5 / 25/ 50 / 100 / 200)
- Time to result: 4.5 hours
- Specificity: Endogenous and recombinant human IL-6
- Protokol booklet
- Validation data file
- Alternative name: Interleukin-6
Clinical significance of Interleukin-6
- Inflammatory diseases: elevated IL-6 levels are detected in various inflammatory conditions such as rheumatoid arthritis (2), systemic lupus erythematosus (3) , and inflammatory bowel disease (4). Therapies targeting Il-6 have been already approved (5).
- Cancer: Il-6 is abundantly present in the tumor microenvironment of various tumours. It promotes tumnorigenesis, invasiveness, and metastases in cancer (6). New perspectives in cancer immunotherapy targeting IL-6 are discussed (7).
- Metabolic diseases: Dysregulated IL-6 activity is associated with pathologies involving chronic inflammation and autoimmunity including metabolic diseases like obesity, metabolic syndrome or type-2 diabetes (8).
RELATED BIOMEDICA KITS
Human VEGF ELISA (BI-VEGF), Human Angiopoietin-2 ELISA (BI-ANG2)
LITERATURE
- Interleukin 6: at the interface of human health and disease. Grebenciucova E, VanHaerents S. Front Immunol. 2023 Sep 28;14:1255533. doi: 10.3389/fimmu.2023.1255533. PMID: 37841263; PMCID: PMC10569068.
- Interleukin-6 in Rheumatoid Arthritis. Pandolfi F, Franza L, Carusi V, Altamura S, Andriollo G, Nucera E. Int J Mol Sci. 2020 Jul 23;21(15):5238. doi: 10.3390/ijms21155238. PMID: 32718086; PMCID: PMC7432115.
- Role of IL-6 and IL-6 targeted therapy in systemic lupus erythematosus. Nepal D, Gazeley D. Rheumatology (Oxford). 2023 Dec 1;62(12):3804-3810. doi: 10.1093/rheumatology/kead416. PMID: 37594751.
- Cytokines of the interleukin-6 family as emerging targets in inflammatory bowel disease. Garbers C, Lokau J. Expert Opin Ther Targets. 2024 Jan-Feb;28(1-2):57-65. doi: 10.1080/14728222.2024.2306341. Epub 2024 Jan 23. PMID: 38217849.
- Therapeutic uses of anti-interleukin-6 receptor antibody. Kang S, Tanaka T, Kishimoto T. Int Immunol. 2015 Jan;27(1):21-9. doi: 10.1093/intimm/dxu081. Epub 2014 Aug 20. PMID: 25142313.
- The Role of IL-6 in Cancer Cell Invasiveness and Metastasis-Overview and Therapeutic Opportunities. Rašková M, Lacina L, Kejík Z, Venhauerová A, Skaličková M, Kolář M, Jakubek M, Rosel D, Smetana K Jr, Brábek J. Cells. 2022 Nov 21;11(22):3698. doi: 10.3390/cells11223698. PMID: 36429126; PMCID: PMC9688109.
- New perspectives in cancer immunotherapy: targeting IL-6 cytokine family. Soler MF, Abaurrea A, Azcoaga P, Araujo AM, Caffarel MM.J Immunother Cancer. 2023 Nov;11(11):e007530. doi: 10.1136/jitc-2023-007530. PMID: 37945321; PMCID: PMC10649711.
- IL-6 family cytokines as potential therapeutic strategies to treat metabolic diseases. Zhao J, Turpin-Nolan S, Febbraio MA. Cytokine. 2021 Aug;144:155549. doi: 10.1016/j.cyto.2021.155549. Epub 2021 May 4. PMID: 33962843.
We are excited to be soon exhibiting at the International Conference on Children’s Bone Health (ICCBH) in Salzburg, Austria from June 22-25, 2024.
Join us at booth number 4 and explore our widely trusted Biomarker ELISA Assay Kits including Fibroblast Growth Factor (FGF23), Sclerostin, Periostin, NT-proCNP, and many others.
We look forward to connecting with you!
Join us at the International Conference on Children’s Bone Health
Click here for more information about the conference for anyone who is interested in
bone metabolism and bone mass in children, adolescents and young adults.
The ICCBH conference aims to unite researchers, clinicians, health professionals, and others from different fields to gain an understanding of the developing skeleton with regards to childhood health and disease. Latest advancements, innovative therapies, and genetic discoveries will be discussed.
More about Biomarkers in Bone Biology
Bone cells release biomarkers during bone remodeling. They can be used in assessing bone diseases and represent useful therapeutic targets. Bone biomarkers can easily be detected in serum and plasma samples by immunoassay.
A recent review by Ponds-Belda OD et al., Mineral Metabolism in Children: Interrelation between Vitamin D and FGF23 focuses on various aspects of FGF23 metabolism in children, reference values, and other key variables involved in mineral homeostasis.
BIOMEDICA offers quality kits to measure various bone biomarkers
Sclerostin ELISA (SOST; cat.no. BI-20492)
OPG ELISA (Osteoprotegerin; cat.no. BI-20403)
RANKL ELISA (soluble RANKL; cat.no. BI-20462)
DKK-1 ELISA (Dickkopf-1; cat.no. BI-20413)
FGF23 intact ELISA (Fibroblast growth factor-23 intact; cat.no. BI-20700)
FGF23 C-terminal ELISA (Fibroblast growth factor-23 C-terminal; cat.no. cat.no. BI-20702)
PERIOSTIN ELISA (POSTN, BI-20433)
NT-proCNP ELISA (NT-C-type natriuretic peptide, BI-20812)
TRUSTED – cited in over 1300 publications
- Kit validations follows international quality guidelines
- Developed & manufactured by Biomedica in Austria
Cardiovascular disease is the leading cause of death in patients with end-stage renal disease on dialysis (1). In an attempt to understand the underlying mechanisms of the high mortality among dialysis patients, other than the traditional cardiovascular risk factors, researchers investigated the association between gut permeability, circulating bacterial fragments, and volume overload in peritoneal dialysis (PD) patients (2).
Gut permeability, circulating bacterial fragments and measures of congestion in peritoneal dialysis.
In this prospective observational study, 108 consecutive adult incident PD patients were recruited and circulating bacterial fragments, N-terminal pro B-type natriuretic peptide (NT-proBNP), calprotectin and zonulin levels were evaluated (2).
NT-proBNP in end-stage kidney disease
Markers of fluid overload
NT-proBNP is recognized as a marker of marker of left ventricular strain and intravascular fluid overload (3, 4).
Fluid overload was quantified by serum levels of NT-proBNP with the Biomedica ELISA kit (Biomedica-cat. no. SK-1204) and with multi-frequency bioimpedance spectroscopy (2).
NT-proBNP ELISA assay kit (cat. no. SK-1204)
√ CE-marked – for IVD use in the EU
√ EASY – simple 2 step protocol, can be run in every lab
√ High and low kit controls included
√ RELIABLE – validated according to international quality guidelines (see validation data )
√ TRUSTED – widely cited in 125 publications
Example of a Biomedica ELISA Assay Kit
Literature
1. Cardiovascular complications in chronic kidney disease: a review from the European Renal and Cardiovascular Medicine Working Group of the European Renal Association. Zoccali C, Mallamaci F, Adamczak M, de Oliveira RB, Massy ZA, Sarafidis P, Agarwal R, Mark PB, Kotanko P, Ferro CJ, Wanner C, Burnier M, Vanholder R, Wiecek A. Cardiovasc Res. 2023 Sep 5;119(11):2017-2032. doi: 10.1093/cvr/cvad083. PMID: 37249051; PMCID: PMC10478756.
2. Gut permeability, circulating bacterial fragments and measures of congestion in peritoneal dialysis. Li C, Ng JK, Chan GC, Fung WW, Lai KB, Poon PY, Luk CC, Chow KM, Szeto CC. Clin Kidney J. 2024 Mar 6;17(3):sfae056. doi: 10.1093/ckj/sfae056. PMID: 38516523; PMCID: PMC10956420.
3. NT-proBNP, fluid volume overload and dialysis modality are independent predictors of mortality in ESRD patients. Paniagua R, Ventura MD, Avila-Díaz M, Hinojosa-Heredia H, Méndez-Durán A, Cueto-Manzano A, Cisneros A, Ramos A, Madonia-Juseino C, Belio-Caro F, García-Contreras F, Trinidad-Ramos P, Vázquez R, Ilabaca B, Alcántara G, Amato D. Nephrol Dial Transplant. 2010 Feb;25(2):551-7. doi: 10.1093/ndt/gfp395. Epub 2009 Aug 12. PMID: 19679559.
4. Determination of volume overload by bioelectrical impedance analysis and NT-proBNP in diabetic pre-dialysis patients. Acta Endocrinol (Buchar). Yildirim Y, Kara AV, Kilinç F, Aydin F, Aydin E, Yilmaz Z, Kadiroglu AK, Yilmaz ME. 2016 Jan-Mar;12(1):19-25. doi: 10.4183/aeb.2016.19. PMID: 31258795; PMCID: PMC6586753.
Proficiency Testing is an external validation of diagnostic methods performed by accredited laboratories (1) as a measure to comply with international quality standards. Through participation in inter-laboratory comparisons such as proficiency testing programs, customers using a diagnostic method (e.g. an ELISA assay), can be assured that results generated from the assay comply with internationally set quality standards.
NT-proBNP assay successfully passes cardiac marker survey
NT-proBNP as a marker of heart failure
Heart failure is a significant cause of morbidity and mortality worldwide. Circulating biomarkers reflecting disease-related pathways that are involved in the development and progression of heart failure (HF) can be helpful in assisting clinicians in the early diagnosis and management of HF patients.
NT-proBNP (N-terminal pro B-type Natriuretic Peptide) is a cardiac protein fragment that is secreted into the bloodstream when the heart muscle stretches (under stress or damage). NT-proBNP is recognized as the hallmark biomarker for heart failure diagnosis and prognosis (2, 3).
NT-proBNP ELISA assay kit (cat. no. SK-1204)
- CE-marked – for IVD use in the EU
- Proficiency tested – click here to access the certificate
- Flexible – can be run in every lab
- Accurate – two controls included
- Easy – 2 step protocol
- Trusted – widely cited in 125 publications
Example of a Biomedica ELISA Assay kit
Developed & manufactured by Biomedica
Literature
- RfB- reference institute for bioanalytics– proficiency testing
- NT-proBNP: The Gold Standard Biomarker in Heart Failure. PM McKie PM and JC Jr Burnett. J Am Coll Cardiol. 2016; Dec 6;68(22):2437-2439. doi: 10.1016/j.jacc.2016.10.001. PMID: 27908348.
- Biomarkers for the diagnosis and management of heart failure. Castiglione V, Aimo A, Vergaro G, Saccaro L, Passino C, Emdin M.Heart Fail Rev. 2022 Mar;27(2):625-643. doi: 10.1007/s10741-021-10105-w. Epub 2021 Apr 14. PMID: 33852110; PMCID: PMC8898236.
Smoking is a major risk factor for a wide range of disorders, including cardiovascular, respiratory diseases, and cancers. Cigarette smoke generates a high level of reactive oxygen species (ROS), causing oxidative stress and cellular damage (1-3). Among the ROS, aldehyds are the main compounds that are implicated in the process of tobacco-induced diseases (3).
Autoantibodies against oxidized-LDL in smokers
Smoking is associated with increased levels of Autoantibodies against oxidized-LDL
In smokers, low density lipoprotein (LDL) is more prone to oxidation due to an excessive presence of reactive ROS, leading to elevated levels of antibodies against oxidized LDL (anti-oxLDL Ab). In healthy individuals no significant differences in anti-oxLDL Ab levels have been observed between smokers and non-smokers. However, moderately elevated levels of anti-oxLDL Ab have been noted in hypercholesterolemic patients (4).
In the same patient group researchers found that foods containing olive oil and lycopene can significantly contribute to reducing LDL-induced oxidative stress (5). The authors suggest that lycopene-olive oil can be used as a supplement to reduce oxidative stress and the inflammatory process in hypercholesterolemic subjects, thereby potentially lowering the risk of atherosclerosis (5).
Function of Low-density lipoprotein (LDL)
Low-density lipoprotein (LDL) is responsible for transporting cholesterol from the liver to various tissues and cells in the body. It is used for essential functions like cell repair, cell membrane structure and hormone production. where it is used fo and other lipids through the bloodstream (6).
Smoking leads to a rise of LDL cholesterol levels, which contributes to plaque buildup in arteries, increasing heart disease and the risk of stroke. Smoking promotes the oxidation of LDL particles, and ox LDL is more likely to adhere to artery walls, leading to atherosclerosis (7).
Antibodies against oxidized LDL (anti-oxLDL Ab).
Oxidized low-densitity lipoprotein (oxLDL) plays an important role in in the development of atherosclerosis. Autoantibodies against oxidatively modified LDL particles can serve as a parameter that consistently reflects the ongoing oxidation processes taking place in vivo. Studies demonstrate increased levels of autoantibodies against oxLDL in the bloodstream of individuals with coronary artery disease (4)
Several studies reported a significant association between anti-oxLDL levels and the subsequent development of Atherosclerosis-related cardiovascular outcomes (4, 8, 9).
Measurement of Antibodies against oxidized LDL (anti-oxLDL Ab)
Autoantibodies targeting oxidatively modified LDL particles can be measured serum with an ELISA assay
Anti-Oxidized LDL Autoantibody ELISA (oLAB) Assay (cat. no. BI-20032)
Features
- Widely cited
- Results in 2,5 hours
- 2 controls included
Literature
- Cigarette Smoke-Induced Reactive Oxygen Species Formation: A Concise Review. Antioxidants (Basel). Seo YS, Park JM, Kim JH, Lee MY. 2023 Sep 7;12(9):1732. doi: 10.3390/antiox12091732. PMID: 37760035; PMCID: PMC10525535.
- Relationships among smoking, oxidative stress, inflammation, macromolecular damage, and cancer. Caliri AW, Tommasi S, Besaratinia A. Mutat Res. 2021 Jan-Jun;787:108365. doi: 10.1016/j.mrrev.2021.108365. Epub 2021 Jan 11. PMID: 34083039; PMCID: PMC8287787.
- Exocyclic DNA adducts and oxidative stress parameters: useful tools for biomonitoring exposure to aldehydes in smokers. Alamil H, Colsoul ML, Heutte N, Van Der Schueren M, Galanti L, Lechevrel M. Biomarkers. 2024 Apr 2:1-7. doi: 10.1080/1354750X.2024.2333361. Epub ahead of print. PMID: 38506499.
- Cigarette smoking potentiates endothelial dysfunction of forearm resistance vessels in patients with hypercholesterolemia. Role of oxidized LDL. Heitzer T, Ylä-Herttuala S, Luoma J, Kurz S, Münzel T, Just H, Olschewski M, Drexler H. Circulation. 1996 Apr 1;93(7):1346-53. doi: 10.1161/01.cir.93.7.1346. PMID: 8641023.
- Beneficial Effects of Olive Oil Enriched with Lycopene on the Plasma Antioxidant and Anti-Inflammatory Profile of Hypercholesterolemic Patients. Martínez Álvarez JR, Lopez Jaen AB, Cavia-Saiz M, Muñiz P, Valls-Belles V. Antioxidants (Basel). 2023 Jul 20;12(7):1458. doi: 10.3390/antiox12071458. PMID: 37507996; PMCID: PMC10376681.
- Biochemistry, Low Density Lipoprotein, Senthil K. Venugopal; McDamian Anoruo; Ishwarlal Jialal. 2024, StatPearls Publishing LLC.
- Smoking and small, dense low-density lipoprotein particles: cross-sectional study. Urahama N, Iguchi G, Shimizu M, Fujihira K, Kobayashi S, Baba H. Nicotine Tob Res. 2008 Aug;10(8):1391-5. doi: 10.1080/14622200802238852. PMID: 18686187.
- Oxidized LDL to autoantibodies against oxLDL ratio – The new biomarker associated with carotid atherosclerosis and cardiovascular complications in dialyzed patients. Pawlak, K., Mysliwiec, M., Pawlak, D., 2012. Atherosclerosis 224, 252–257.
- Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou, E., Vourli, G., Nounopoulos, C., Zachari, A., Dionyssiou-Asteriou, A., 2005. Free Radic. Res. 39, 419–429.
Rheumatoid arthritis is a chronic inflammatory disorder that affects not only the joints but can also damage other parts of the body including the skin, lungs (1), heart and blood vessels. Patients with RA have a notable 50-70% increased risk of developing cardiovascular diseases compared to the general population (2).
Effects of Tofacitinib Therapy on Angiogenic Biomarkers in Rheumatoid Arthritis
In a recent study, highlighting the Biomedica NT-proBNP ELISA Kit*, researchers investigated for the first time the effects of 1-year Tofacitinib therapy on angiogenic biomarkers in relation to vascular inflammation and function as well as clinical markers in patients with rheumatoid arthritis (RA) (3). Tofacitinib is a medication used to treat RA. It inhibits Janus kinase enzymes which are involved in cytokine signaling leading to inflammation and symptoms of RA (4).
*Serum NT-proBNP (pmol/l) concentrations were detected by commercially available ELISA kits (NT-proBNP ELISA, Biomedica, Vienna, Austria)
NT-proBNP ELISA Kit (cat. no. SK-1204)
- Quality – CE marked – for IVD use in the EU (and proficiency tested*)
- Easy – simple protocol, kit includes two controls
- Reliable – fully validated
- Trusted – Cited in 125 publications
Download the Biomedica NT-proBNP ELISA Assay Proficiency Testing Certificate here
About Proficiency Testing
Proficiency Testing offers a report enabling laboratories to benchmark their data against data from other laboratories globally. Conducted within a ring trial program, this testing is executed by accredited laboratories specialized on proficiency testing. Proficiency testing oversees the performance of individual laboratories for specific tests like cardiac biomarker assays.
Abstract
Objectives – Cardiovascular (CV) morbidity and mortality, and perpetuated synovial angiogenesis have been associated with RA. In our study we evaluated angiogenic factors in relation to vascular inflammation and function, and clinical markers in RA patients undergoing 1-year tofacitinib therapy.
Methods – Thirty RA patients treated with either 5 mg or 10 mg twice daily tofacitinib were included in a 12-month follow-up study. Eventually, 26 patients completed the study and were included in data analysis. Levels of various angiogenic cytokines (TNF-α, IL-6), growth factors [VEGF, basic fibroblast (bFGF), epidermal (EGF), placental (PlGF)], cathepsin K (CathK), CXC chemokine ligand 8 (CXCL8), galectin-3 (Gal-3) and N-terminal prohormone brain natriuretic peptide (NT-proBNP) were determined at baseline, and at 6 and 12 months after initiating tofacitinib treatment. In order to assess flow-mediated vasodilation, common carotid intima-media thickness (ccIMT) and carotid-femoral pulse-wave velocity, ultrasonography was performed. Synovial and aortic inflammation was also assessed by 18F-fluorodeoxyglucose-PET/CT.
Results – One-year tofacitinib therapy significantly decreased IL-6, VEGF, bFGF, EGF, PlGF and CathK, while it increased Gal-3 production (P < 0.05). bFGF, PlGF and NT-proBNP levels were higher, while platelet-endothelial cell adhesion molecule 1 (PECAM-1) levels were lower in RF-seropositive patients (P < 0.05). TNF-α, bFGF and PlGF correlated with post-treatment synovial inflammation, while aortic inflammation was rather dependent on IL-6 and PECAM-1 as determined by PET/CT (P < 0.05). In the correlation analyses, NT-proBNP, CXCL8 and Cath variables correlated with ccIMT (P < 0.05).
Conclusions – Decreasing production of bFGF, PlGF or IL-6 by 1-year tofacitinib therapy potentially inhibits synovial and aortic inflammation. Although NT-proBNP, CXCL8 and CathK were associated with ccIMT, their role in RA-associated atherosclerosis needs to be further evaluated.
Literature
- Identification, Monitoring, and Management of Rheumatoid Arthritis-Associated Interstitial Lung Disease. Koduri G and Solomon JJ. Arthritis Rheumatol, 2023; Dec;75(12):2067-2077. doi: 10.1002/art.42640.
- Association of cardiovascular risks in rheumatoid arthritis patients: Management, treatment and future perspectives. Nishant Johri et al., 2023; Health Sciences Review, Vol 8.
- Effects of 1-year tofacitinib therapy on angiogenic biomarkers in rheumatoid arthritis. Kerekes G et al., Rheumatology (Oxford), 2023; 62(SI3):SI304-SI312. doi: 10.1093/rheumatology/kead502.
- Efficacy and safety of JAK inhibitors in rheumatoid arthritis: update for the practising clinician. Szekanecz Z et al., Nat Rev Rheumatol, 2024; 20(2):101-115. doi: 10.1038/s41584-023-01062-9.
Lyme disease (LD) or Lyme Borreliosis is a bacterial infection that can be transmitted to humans by infected ticks. It is estimated that about 700 000 individuals in Europe and in the United States are infected with LD every year, though the number of unrecorded or misdiagnosed cases could be higher (1).
Symptoms include fever, headache, joint pain, and swollen lymph nodes. Lyme disease is often accompanied by a distinctive expanding rash (Erythema Migrans) which starts at the site of the tick bite (2). In most cases when LD is diagnosed early, treatment with antibiotics is very effective. However, without treatment, the infection can spread to different areas of the body, impacting the joints, heart, and nervous system (3). Early diagnosis and treatment are essential for managing Lyme disease effectively.
The Biomedica Borrelia ELISA Assays were utilized in a recent study to estimate the incidence of symtopmatic Lyme Borreliosis cases in Lubin, Poland in 2021: Estimated Incidence of Symptomatic Lyme Borreliosis Cases in Lublin, Poland in 2021. Colby E et al., Microorganisms. 2023; 11(10):2481. The authors concluded that “after adjusting for under-ascertainment, the estimated number of symptomatic LB cases in Lublin in 2021 was 6204 (population-based incidence: 467.6/100,000). After adjustment for under-ascertainment, the incidence of symptomatic LB in Lublin, Poland, is high.”
Serological Testing for Lyme Disease with BIOMEDICA ELISA Kits
The Borrelia assays from Biomedica use recombinant antigens to specifically detect IgM and IgG antibodies against the immunodominant antigens of the three genospecies (Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii) and thus offer:
- High sensitivity and specificity that is confirmed by clinical samples
- Standardized CSF and serum analysis available
- For manual and automated testing
- No extra RF stripping necessary
- Widely cited
- CE marked – for IVD use in the EU
Borrelia IgM ELISA (cat. no. BI-21042)
ENZYME IMMUNOASSAY FOR THE QUALITATIVE OR QUANTITATIVE DETERMINATION OF IgG ANTIBODIES TO BORRELIA IN PLASMA, SERUM OR CEREBROSPINAL FLUID
The following recombinant antigens are utilized in the Biomedica Borrelia IgM ELISA Assay:
- p21 OspC – B. afzellii (pKo)
- p21 OspC – B. garinii (20047)
- p41/I – B. bavariensis (pBi)
- VIsE – fusion proteins of different Borrelia genospecies
Borrelia IgG ELISA (cat. no. BI- 21032)
ENZYME IMMUNOASSAY FOR THE QUALITATIVE OR QUANTITATIVE DETERMINATION OF IgG ANTIBODIES TO BORRELIA IN PLASMA, SERUM OR CEREBROSPINAL FLUID
The following recombinant antigens are utilized in the Biomedica Borrelia IgG ELISA Assay:
- p21 – OspC – B. burgdorferi sensu stricto (B31), B. garinii (20047)
- p18 – B. afzelii (pKo)
- p100 – B. afzelii (pKo)
- VIsE – fusion protein of different Borrelia genospecies
Literature
- Lyme borreliosis diagnosis: state of the art of improvements and innovations. Guérin M et al., 2023; BMC Microbiol, 23(1):204.
- Lyme borreliosis. Steere AC et al., 2016 ; Rev Dis Primers. 2:16090.
- Neurologic manifestations of Lyme Borreliosis. Summer G et al., 2019; Rev Neurol (Paris). 175(7-8):417-419.
The Biomedica soluble SEMAPHORIN 4D Assay Kit (sSEMA4D) was utilized in a recent study assessing the role of sSEMA4D in critically ill and septic patients as a biomarker for prediction and disease severity or survival (1). The results revealed that blood SEMA4D concentrations are increased in patients with liver cirrhosis and are positively correlated with liver function tests. The study suggests that sSEMA4D may be associated with hepatic injury and inflammation and may serve as a potential biomarker in critically ill patients with liver cirrhosis.
Semaphorin 4D in critically ill patients
About Semaphorin 4D
Semaphorin 4D (Sema4D) also known as CD100 is a member of the Sema4D family with established immunregulatory functions (2). Sema 4D exists as a membrane bound and in a soluble form. It is expressed on immune cells and is upregulated upon cellular activation resulting in shedding of its extracellular soluble Sema4D domain. This soluble biologically active form has a molecular weight of 120KD and can be measured in blood samples (3, 4). Due to its regulatory role in the immune system the potential application of Sema4D as a diagnostic marker and as a therapeutic target for the treatment of immunological disorders is currently under investigation (5).
Biomedica soluble SEMAPHORIN 4D (sSEMA4D) ELISA Assay Kit (cat. no. BI-20405)
Features & Benefits
- Only Sema4D ELISA assay that is fully validated
- 10 µl / well sample volume – no sample predilution
- Reference values provided
Links to soluble Semaphorin 4D ELISA – protocol booklet – validation data file
Literature
- Soluble Semaphorin 4D Serum Concentrations Are Elevated in Critically Ill Patients with Liver Cirrhosis and Correlate with Aminotransferases. Abu Jhaisha S, Hohlstein P, Yagmur E, Köller V, Pollmanns MR, Adams JK, Wirtz TH, Brozat JF, Bündgens L, Hamesch K, Weiskirchen R, Tacke F, Trautwein C, Koch A. Diagnostics (Basel). 2024 Feb 8;14(4):370. doi: 10.3390/diagnostics14040370. PMID: 38396409; PMCID: PMC10887520.
- The Role of Semaphorin 4D in Bone Remodeling and Cancer Metastasis. Lontos K, Adamik J, Tsagianni A, Galson DL, Chirgwin JM, Suvannasankha A. Front Endocrinol (Lausanne). 2018 Jun 19;9:322. doi: 10.3389/fendo.2018.00322. PMID: 29971044; PMCID: PMC6018527.
- Soluble SEMA4D/CD100: A novel immunoregulator in infectious and inflammatory diseases. Maleki KT, Cornillet M, Björkström NK. Clin Immunol. 2016 Feb;163:52-9. doi: 10.1016/j.clim.2015.12.012. Epub 2015 Dec 28. PMID: 26732857.
- A high-sensitivity enzyme immunoassay for the quantification of soluble human semaphorin 4D in plasma. Laber A, Gadermaier E, Wallwitz J, Berg G, Himmler G. Anal Biochem. 2019 Jun 1;574:15-22. doi: 10.1016/j.ab.2019.03.004. Epub 2019 Mar 14. PMID: 30879960.
- The role of Sema4D/CD100 as a therapeutic target for tumor microenvironments and for autoimmune, neuroimmune and bone diseases. Wu M, Li J, Gao Q, Ye F. Expert Opin Ther Targets. 2016 Jul;20(7):885-901. doi: 10.1517/14728222.2016.1139083. Epub 2016 Jan 28. PMID: 26732941.
Irritable bowel syndrome (IBS) is a common gastrointestinal disorder affecting approximately around 1 in 10 people globally (1).
IBS is characterized by recurrent abdominal pain and changes in bowel habits. The exact causes of IBS is unknown but stress, gastrointestinal infections and a family history of IBS may play a role (1,2). IBS is usually a lifelong problem and symptoms have an effect on daily life. Managing symptoms through diet and lifestyle changes can improve the quality of life (2, 3, 4) .
Accepted biomarkers for IBS are still missing. Recent studies have shown that Leucine-rich alpha glycoprotein (LRG) may be a novel marker to evaluate disease activity and mucosal healing in IBS (5, 6). Moreover, researchers have recently proposed that serum LRG may be a potential diagnostic marker for pediatric IBD (7).
Biomarkers for Irritable Bowel Syndrome
Leucine-rich alpha glycoprotein (LRG)
Leucine-rich alpha-2 glycoprotein (LRG) (also named LRG1) has a molecular weight of 50 kDa glycoprotein and contains repetitive sequences with a leucine-rich motif. LRG derives predominantly from immune cells i.e. neutrophils, macrophags but is also secreted by intestinal epithelial cells and hepatocytes (8) in response to various cytokines (TNF-alpha, IL-6 and IL-22).
LRG has been identified as an inflammatory biomarker for various immune-mediated diseases including IBD, rheumatoid arthritis, juvenile idiopathic arthritis and others (8).
LRG can be measured in human serum and plasma samples by ELISA Assay
LRG ELISA (cat. no. BI-LRG) – developed and manufactured by BIOMEDICA
- RELIABLE – validated following international quality guidelines (FDA, EMA, ICH) . The validation data can be found here.
- REFERENCE values provided for normal and pathological samples
- OPTIMIZED – assay range optimized for clinical samples, no additional testing required
- EASY – results in 3.5 h (protocol booklet)
- CONVENIENT – all reagents included; 7 pre-diluted standards/calibrators, 2 controls and sufficient assay buffer
Example of a Biomedica ELISA Assay Kit
Literature
- Global burden of irritable bowel syndrome: trends, predictions and risk factors. Black CJ et al., Nat Rev Gastroenterol Hepatol. 2020; 17(8):473-486. PMID: 32296140.
- Irritable bowel syndrome. Ford AC et al., Lancet. 2020; 396(10263):1675-1688. PMID: 33049223.
- A Discussion of Whether Various Lifestyle Changes can Alleviate the Symptoms of Irritable Bowel Syndrome. Okawa Y. Healthcare (Basel). 2022; 10(10):2011. PMID: 36292457.
- Irritable bowel syndrome: treatment based on pathophysiology and biomarkers. Camilleri M, Boeckxstaens G. Gut. 2023;72(3):590-599. PMID: 36307180.
- Leucine-rich alpha-2 glycoprotein as a marker of mucosal healing in inflammatory bowel disease. Yasutomi E et al., Sci Rep. 2021; 27;11(1):11086.
- Leucine-Rich Alpha-2 Glycoprotein Is a Reliable Serum Biomarker for Evaluating Clinical and Endoscopic Disease Activity in Inflammatory Bowel Disease. Shimoyama Tet al., Inflamm Bowel Dis. 2023; 29(9):1399-1408.
- Significance of Serum Leucine-rich Alpha-2 Glycoprotein as a Diagnostic Marker in Pediatric Inflammatory Bowel Disease. Yoshimura S et al., Kobe J Med Sci. 2024; 69(4):E122-E128. PMID: 38379274.
- Evaluation of Serum Leucine-Rich Alpha-2 Glycoprotein as a New Inflammatory Biomarker of Inflammatory Bowel Disease. Yoshimura Tet a., 2021; 2021:8825374. PMID: 33623482.
Traumatic brain injury (TBI) is a sudden injury that causes damages to the brain. It is one of the most common causes of disability and death in adults (1). There are currently no early biomarkers for prognosis in routine clinical use (2). Apart from the initial injury, victims with TBI face the possibility of secondary neurological damage. Studies have identified TBI as a risk factor for all-cause dementia and Parkinson’s disease (3).
Interleukin-6 in Traumatic Brain Injury
Interleukin (IL)-6 is a proinflammatory cytokine that plays a key role in the immune response to acute neurological injury (1, 4) . Studies have shown that IL-6 may serve as a prognostic biomarker of clinical outcomes after traumatic brain injury (2).
IL-6 can reliably be measured with the highly sensitive IL-6 ELISA assay from Biomedica
IL-6 ELISA (Biomedica, cat. no. BI-IL6)
- HIGHLY SENSITIVE- measurable values in serum and plasma samples
- EASY – ready to use calibrators and controls
- RELIABLE – full validation package
Literature
- Interleukin-6 in Traumatic Brain Injury: A Janus-Faced Player in Damage and Repair. Ciryam P, Gerzanich V, Simard JM. J Neurotrauma. 2023 Nov;40(21-22):2249-2269. doi: 10.1089/neu.2023.0135. Epub 2023 Aug 10. PMID: 37166354; PMCID: PMC10649197.
- Interleukin-6 as a prognostic biomarker of clinical outcomes after traumatic brain injury: a systematic review. Ooi SZY, Spencer RJ, Hodgson M, Mehta S, Phillips NL, Preest G, Manivannan S, Wise MP, Galea J, Zaben M. Neurosurg Rev. 2022 Oct;45(5):3035-3054. doi: 10.1007/s10143-022-01827-y. Epub 2022 Jul 6. PMID: 35790656; PMCID: PMC9256073.
- Traumatic Brain Injury and Risk of Neurodegenerative Disorder. Brett BL, Gardner RC, Godbout J, Dams-O’Connor K, Keene CD. Biol Psychiatry. 2022 Mar 1;91(5):498-507. doi: 10.1016/j.biopsych.2021.05.025. Epub 2021 Jun 2. PMID: 34364650; PMCID: PMC8636548.
- Role of IL-6 in the regulation of neuronal development, survival and function. Kummer KK, Zeidler M, Kalpachidou T, Kress M. Cytokine. 2021 Aug;144:155582. doi: 10.1016/j.cyto.2021.155582. Epub 2021 May 29. PMID: 34058569.
Testing Cytotoxic Activity of Drug Candidates with EZ4U – MTT Assay
Globally, 19.3 million new cancer cases and nearly 10.0 million cancer deaths have been reported in 2020 (1). Despite the significant advances in cancer treatment, several challenges still remain e.g. avoiding resistance or cytotoxic effects. In search of more tolerable and effective drugs, a group of drug candidates have emerged in cancer research that target unique cell death mechanisms.
Ferroptosis is a form of regulated iron-dependent type of cell death that is driven by lipid peroxidation. It offers a new approach for developing novel drugs for cancer treatment. In the last years, several novel compounds capable of inducing ferroptosis in cancer have been investigated. Bernkop-Schnürch et al., recently evaluated the inhibitory effects of different methoxylated complexes on proliferation, migration, and metabolic activity in human breast cancer and leukemia cells. The researchers identified one promising lead compound for the design of a drug candidate that is capable of inducing ferroptosis in cancer cells (2). Read more: Design, Synthesis, Electrochemical, and Biological Evaluation of Fluorescent Chlorido[N,N‘-bis(methoxy/hydroxy)salicylidene-1,2-bis(4-methoxyphenyl)ethylenediamine]iron(III) Complexes as Anticancer Agents.
The Biomedica EZ4U Cell Proliferation and Cytotoxicity Assay (cat. no. BI-5000) was used in this study to evaluate the cytotoxic activity of the novel compounds. The assay is a modified MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) that detects the reduction of tetrazolium salts into yellow-colored formazan derivatives by functional mitochondria.
Brief, the metabolic activity was analysed by seeding cells in microtiter plates and culturing them at 37°C for 3 days in a 5% CO2/95% air atmosphere. Thereafter, 20µl of the EZ4U substance was added to each well and the color change was measured on a microtiter plate reader at a wavelength of 450 nm with 620 nm as a reference.
EZ4U Cell Viability& Cytotoxicity Assay (cat.no. BI-5000)
- SAFE – non-radioactive & non-toxic
- CONVENIENT – single-step incubation for use on living cells
- TRUSTED – widely cited in over 260 publications
Download the BROCHURE – EZ4U cell proliferation and cytotoxicity assay and the PROTOCOL BOOKLET
Literature
- Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F.CA Cancer J Clin. 2021 May;71(3):209-249. doi: 10.3322/caac.21660. Epub 2021 Feb 4. PMID: 33538338.
- Design, Synthesis, Electrochemical, and Biological Evaluation of Fluorescent Chlorido[N,N‘-bis(methoxy/hydroxy)salicylidene-1,2-bis(4-methoxyphenyl)ethylenediamine]iron(III) Complexes as Anticancer Agents. Bernkop-Schnürch AD, Chavooshi D, Descher HA, Leitner D, Talasz H, Hermann M, Wurst K, Hohloch S, Gust R, Kircher B. J Med Chem. 2023 Dec 14;66(23):15916-15925. doi: 10.1021/acs.jmedchem.3c01359. Epub 2023.
Machine Learning for Bone Biomarker Profiling in Rheumatoid Arthritis
Rheumatoid arthritis (RA) is a chronic, progressive inflammatory disorder which can lead to severe joint damage and disability. In 2019, an estimated 18 million people worldwide were living with this disease (1). Untreated RA can lead to destruction of the joints as well as heart, lung or nervous system problems (2). Skeletal bone loss, referred to as osteopenia or osteoporosis, is a key feature of RA.
Sclerostin and Dickkopf-1 (DKK-1) are Wnt signaling proteins that are secreted by osteocytes, bone cells embedded in the bone matrix. They are inhibitors of bone formation and play a key role in the pathogeneses of systemic and localized bone loss in RA (3, 4). Serum levels of Sclerostin and DKK-1 have shown to be elevated in patients with RA compared to controls and correlate with bone erosions and inflammation (4, 5, 6). Findings in mice have demonstrated that DKK-1 triggers inflammatory bone degradation and neutralization of DKK-1 protects from systemic bone loss during inflammation (7). Interestingly, blocking the bone destruction molecule Sclerostin with an anti-sclerostin antibody has shown to be effective for the treatment of osteoporosis but may not be safe for patients suffering from inflammatory RA: in a rodent RA model, Weymeyer et al. demonstrated that Sclerostin inhibition did not stop bone loss and worsened clinical RA outcome by promoting TNF-dependent inflammatory joint destruction (8).
Controlling inflammation by biological therapies targeting pro-inflammatory cytokines has shown to have a positive effect in RA patients (5). Interleukin-6 is a key immunomodulatory cytokine that plays an important role in the development of RA. Inhibition of IL-6 has proven to be effective in treating patients with RA (9). A study by Briot et al showed that DKK-1 levels decreased in RA patients treated with an anti-IL-6 inhibitor (6).
Machine Learning for Bone Biomarker Profiling in Rheumatoid Arthritis
A recent cross-sectional study by Adami G et al., with over 1800 enrolled participants diagnosed with RA, Psoriatic Arthritis (PsA), and Systemic Sclerosis (SSc), employed machine learning techniques to assess the capability of biomarker profiles in differentiating RA patients from individuals with PsA and SSc. The Wnt signaling antagonists Sclerostin and Dickkopf-1 (DKK-1) were among the biomarkers measured. The study provided an in-depth understanding into the bone signature of RA that is marked by changes in bone mineral density and by unique biomarker profiles (6). Serum Sclerostin and DKK-1 levels were measured with ELISA assay kits from Biomedica.
SCLEROSTIN ELISA (#BI-20492) and DKK-1 (#BI-20403) ELISA kits
Biomedica´s Sclerostin ELISA Assay
- TRUSTED – cited in more than 290 publications!
- QUALITY – validated according to international guidelines
- EFFICIENT – only 20µl sample / well
- CONVENIENT – ready to use standards and controls included
- TRUSTED – cited in more than 180 publications!
- QUALITY – validated according to international guidelines
- EFFICIENT – only 20µl sample / well
- CONVENIENT – direct measurement – no sample pre-dilution. Ready to use standards and controls included
Also available from Biomedica : Bioactive Sclerostin ELISA (cat. no. BI-20472), Interleukin-6 ELISA (BI-IL6)
Complete ready-to use ELISA kits
Literature
- GBD 2019: Global burden of 369 diseases and injuries in 204 countries and territories, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019. https://vizhub.healthdata.org/gbd-results
- WHO- Rheumatoid arthritis, June 2023
- Wnt Signaling and Biological Therapy in Rheumatoid Arthritis and Spondyloarthritis. Cici D, Corrado A, Rotondo C, Cantatore FP. Int J Mol Sci. 2019 Nov 7;20(22):5552. doi: 10.3390/ijms20225552. PMID: 31703281; PMCID: PMC6888549.
- Study of correlation of level of expression of Wnt signaling pathway inhibitors sclerostin and dickkopf-1 with disease activity and severity in rheumatoid arthritis patients. Singh A, Gupta MK, Mishra SP.Drug Discov Ther. 2019;13(1):22-27. doi: 10.5582/ddt.2019.01011. PMID: 30880318.
- The effect of tocilizumab on bone mineral density, serum levels of Dickkopf-1 and bone remodeling markers in patients with rheumatoid arthritis. Briot K, Rouanet S, Schaeverbeke T, Etchepare F, Gaudin P, Perdriger A, Vray M, Steinberg G, Roux C. Joint Bone Spine. 2015 Mar;82(2):109-15. doi: 10.1016/j.jbspin.2014.10.015. Epub 2014 Dec 31. PMID: 25557658.
- Machine learning to characterize bone biomarkers profile in rheumatoid arthritis. Adami G, Fassio A, Rossini M, Benini C, Bixio R, Rotta D, Viapiana O, Gatti D. Front Immunol. 2023 Nov 9;14:1291727. doi: 10.3389/fimmu.2023.1291727. PMID: 38022514; PMCID: PMC10665911.
- Neutralisation of Dkk-1 protects from systemic bone loss during inflammation and reduces sclerostin expression. Heiland GR, Zwerina K, Baum W, Kireva T, Distler JH, Grisanti M, Asuncion F, Li X, Ominsky M, Richards W, Schett G, Zwerina J. Ann Rheum Dis. 2010 Dec;69(12):2152-9. doi: 10.1136/ard.2010.132852. Epub 2010 Sep 21. PMID: 20858621.
- Sclerostin inhibition promotes TNF-dependent inflammatory joint destruction. Wehmeyer C, Frank S, Beckmann D, Böttcher M, Cromme C, König U, Fennen M, Held A, Paruzel P, Hartmann C, Stratis A, Korb-Pap A, Kamradt T, Kramer I, van den Berg W, Kneissel M, Pap T, Dankbar B. Sci Transl Med. 2016 Mar 16;8(330):330ra35. doi: 10.1126/scitranslmed.aac4351. Epub 2016 Mar 16. PMID: 27089204.
- Targeting IL-6 or IL-6 Receptor in Rheumatoid Arthritis: What Have We Learned? Avci AB, Feist E, Burmester GR. BioDrugs. 2024 Jan;38(1):61-71. doi: 10.1007/s40259-023-00634-1. Epub 2023 Nov 21. PMID: 37989892; PMCID: PMC10789669.
The Biomedica SCLEROSTIN ELISA Assay Kit (# BI-20492) was utilized in a recent publication assessing the associations between serum and bone sclerostin levels and biomarkers of bone turnover and bone histomorphometry. Read more: Sclerostin, Osteocytes, and Wnt Signaling in Pediatric Renal Osteodystrophy.
Sclerostin (SOST) ELISA (cat. no. BI-20492)
- Most referenced Sclerostin ELISA in over 290 citations
- Low sample volume – 20µl / well
- Validation following international guidelines
Example of a Biomedica ELISA assay kit
Sclerostin a biomarker in renal pediatric bone disease
Sclerostin, Osteocytes, and Wnt Signaling in Pediatric Renal Osteodystrophy. Laster M. et al., Nutrients. 2023 Sep 25;15(19):4127. doi: 10.3390/nu15194127. PMID: 37836411; PMCID: PMC10574198 . link to full text
Abstract
The pathophysiology of chronic kidney disease-mineral and bone disorder (CKD-MBD) is not well understood. Specific factors secreted by osteocytes are elevated in the serum of adults and pediatric patients with CKD-MBD, including FGF-23 and sclerostin, a known inhibitor of the Wnt signaling pathway. The molecular mechanisms that promote bone disease during the progression of CKD are incompletely understood. In this study, we performed a cross-sectional analysis of 87 pediatric patients with pre-dialysis CKD and post-dialysis (CKD 5D). We assessed the associations between serum and bone sclerostin levels and biomarkers of bone turnover and bone histomorphometry. We report that serum sclerostin levels were elevated in both early and late CKD. Higher circulating and bone sclerostin levels were associated with histomorphometric parameters of bone turnover and mineralization. Immunofluorescence analyses of bone biopsies evaluated osteocyte staining of antibodies towards the canonical Wnt target, β-catenin, in the phosphorylated (inhibited) or unphosphorylated (active) forms. Bone sclerostin was found to be colocalized with phosphorylated β-catenin, which suggests that Wnt signaling was inhibited. In patients with low serum sclerostin levels, increased unphosphorylated “active” β-catenin staining was observed in osteocytes. These data provide new mechanistic insight into the pathogenesis of CKD-MBD and suggest that sclerostin may offer a potential biomarker or therapeutic target in pediatric renal osteodystrophy.
Related Literature
FGF-23 and sclerostin in serum and bone of CKD patients. Lima F, Monier-Faugere MC, Mawad H, David V, Malluche HH. Clin Nephrol. 2023 May;99(5):209-218. doi: 10.5414/CN111111. PMID: 36970967; PMCID: PMC10286735. (Biomedica Sclerostin ELISA Assay Kit, cat. no. BI-20492 citation)
Sclerostin and Dickkopf-1 in renal osteodystrophy. Cejka D, Herberth J, Branscum AJ, Fardo DW, Monier-Faugere MC, Diarra D, Haas M, Malluche HH. Clin J Am Soc Nephrol. 2011 Apr;6(4):877-82. doi: 10.2215/CJN.06550810. Epub 2010 Dec 16. PMID: 21164019; PMCID: PMC3069382. (Biomedica Sclerostin ELISA Assay Kit, cat. no. BI-20492 citation)
Bone Disorders in Pediatric Chronic Kidney Disease: A Literature Review. Capossela L, Ferretti S, D’Alonzo S, Di Sarno L, Pansini V, Curatola A, Chiaretti A, Gatto A.Biology (Basel). 2023 Nov 2;12(11):1395. doi: 10.3390/biology12111395. PMID: 37997994; PMCID: PMC10669025.
Biomarkers have a longstanding role as indicators of biological changes. In drug development they serve dual purposes: predicting drug efficacy and identifying potential drug toxicity. Cardiotoxicity in one of the causes why preclinical safety tests fail during drug development. Therefore, monitoring cardiac toxicity with biomarkers is an essential part during drug development.
Cardiac Safety Biomarkers for Preclinical Cardiotoxicity Testing
The cardiac biomarkers NT-proANP and NT-proBNP
The biomarkers NT-proANP and NT-proBNP are cardiac hormones released in response to the stretching of the heart muscle . They belong to the group of natriuretic peptides and serve as cardiac biomarkers in both human and preclinical settings (1-4).
Biomedica offers reliable ELISA kits for measuring NT-proBNP and NT-proANP in both human and rodent samples.
Example of a Biomedica ELISA kit
Rat NT-proBNP ELISA – NEW
- Product code: BI-1204R
- Sample type: rat serum, plasma
- Sample volume: 10 µL/well
- Sensitivity: LOD 21 pg/ml
- Standard curve range: 0 – 3,200 pg/ml
- Specificity: Endogenous (natural) and recombinant human NT-proBNP (1-76)
- Assay time: 3.5 hours
- Reference value information for rat samples is provided. Instructions for use
- Citation click here (5)
also available: NT-proBNP ELISA (cat.no. SK-1204) – for human serum and plasma samples
- CE marked for IVD use in the EU
- Proficiency testing , saliva protocol
- Widely cited
- Product code: BI-20892
- Sample types: Serum, plasma, urine, cell culture supernatant (human, rat, mouse, rabbit samples)
Due to the high sequence homology of NT-proANP among species the assay has been used not only in human samples but also in rodent (rat, mouse) rabbit, dog samples
- Sample volume: 10 µL/well
- Sensitivity: LOD 0.05 nmol/l (= 0.64 ng/ml)
- Standard curve range: 0 – 10 nmol/l (= 0 – 127 ng/ml)
- Specificity: endogenous (natural) and recombinant human NT-proANP (equivalent to proANP 1-98)
- There is high cross-reactivity between animal species.
- Assay time: 3.5 hours
- Citations all , citations with use of rat/mouse samples
- Instructions for use
NT-proANP – a marker of drug-induced hypertrophy in rats
The validity of NT-proANP measurements as a tool for the detection of cardiovascular disorders in rats was demonstrated in an interlaboratory comparative study using Biomedica’s NT-proANP ELISA assay (Cat. No. BI-20892). Rat samples were analyzed and evaluated among 5 different laboratories by the PSTC-CHWG (Predictive Safety Testing Consortium and Cardiac Hypertrophy Working Group). The results demonstrated that the Biomedica NT-proANP ELISA assay (#BI-20892) is robust and technically adequate for the detection of serum NT-proANP levels in SD rats (6, 7).
Literature
- Atrial and brain natriuretic peptides: Hormones secreted from the heart. Nakagawa Y, Nishikimi T, Kuwahara K. Peptides. 2019 Jan;111:18-25. doi: 10.1016/j.peptides.2018.05.012. Epub 2018 May 31. PMID: 29859763.
- Cardiac natriuretic peptides. Goetze JP, Bruneau BG, Ramos HR, Ogawa T, de Bold MK, de Bold AJ. Nat Rev Cardiol. 2020 Nov;17(11):698-717. doi: 10.1038/s41569-020-0381-0. Epub 2020 May 22. PMID: 32444692.
- Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy . Dunn, M.E., Manfredi, T.G., Agostinucci, K., Engle, S.K., Powe J., King, N.M.P., Rodriguez, L.A, Gropp, K.E., Gallacher, M., Vetter, F.J., More, V., Shimpi, P., Serra, D., Colton, H.M., for The Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium . 2017. Toxicol Pathol 45(2): 334-352. PMID: 27102652
- Natriuretic Peptides as Cardiovascular Safety Biomarkers in Rats: Comparison With Blood Pressure, Heart Rate, and Heart Weight . Engle, S.K., and Watson, D.E. 2016. Toxicol Sci 149: 458 – 472. PMID: 26609138.
- Leucine Supplementation Improves Diastolic Function in HFpEF by HDAC4 Inhibition. Alves PKN, Schauer A, Augstein A, Männel A, Barthel P, Joachim D, Friedrich J, Prieto ME, Moriscot AS, Linke A, Adams V Cells. 2023 Nov 2;12(21):2561. doi: 10.3390/cells12212561. PMID: 37947639; PMCID: PMC10648219.
- Cross-laboratory analytical validation of the cardiac biomarker NT-proANP in rat. Vinken P, Reagan WJ, Rodriguez LA, Buck WR, Lai-Zhang J, Goeminne N, Barbacci G, Liu R, King NM, Engle SK, Colton H.J Pharmacol Toxicol Methods. 2016; 77:58-65. doi: 10.1016/j.vascn.2015.10.002. PMID: 26516096.
- Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy. Dunn ME et al., Toxicol Pathol. 2017; 45(2):344-352.
Kidney disease is estimated to affect more than 800 million people worldwide (1). Taking action for prevention, early diagnosis and treatment can reduce the risk and the burden of kidney disease.
Role of the kidneys, kidney disease and risk factors
Kidneys are vital organs that regulate fluid balance, blood pressure and produce hormones that stimulate the production of red blood cells . Kidney disease is a condition in which kidneys lose their ability to effectively filter waste products and excess fluids from the blood. Kidney disease commonly leads to a decline in kidney function that may lead to kidney failure, characterized by the complete loss of kidney function. At this stage dialysis or kidney transplantation become the only treatment option.
Kidney problems can emerge suddenly (acute) or gradually over time (chronic). Various conditions, diseases and medications can contribute to acute and chronic kidney problems. Chronic kidney disease (CKD) is characterized by a prolonged period of kidney abnormalities that last for more than three months (2), whereas acute kidney disease – acute kidney injury (AKI) is characterized by a sudden loss of excretory kidney function (3).
Other forms of kidney disease include polycystic kidney disease (PKD) a genetic disorder that leads to kidney enlargement and impaired kidney function over time and glomerulonephritis (GN). GN is a group of diseases characterized by inflammation of the glomeruli, the filtration units of the kidney (4).
The most common risk factors for the development and progression of CKD are diabetes and high blood pressure. Managing blood sugar and blood pressure can help keep kidneys healthy. Other risk factors of CKD include heart disease, obesity, family history and genetic background as well as age, smoking and nutrition (5).
Consequences of kidney disease include heart disease, high blood pressure, bone and mineral disorders, and anemia (6).
Keep Your Kidneys Healthy
Regular exercise, weight control, a balanced diet (7) and sufficient fluid intake are only some of the ways to keep your kidney healthy.
World Kidney Day – about kidney health download here and check out the 6-Step Guide to Protecting Kidney Health here .
BIOMEDICA – Biomarker ELISA kits for clinical research in kidney disease
- HIGH QUALITY ASSAYS – Fully validated according to international quality guidelines
- TRUSTED – Widely cited in over 1500 publications
ELISA kits for: FGF23 (Fibroblast growth factor 23), Vanin-1, Endostatin, Sclerostin, Osteoprotegerin (OPG), Angiopoietin-2 (ANG-2), IL-6 (Interleukin-6) , VEGF (Vascular Endothelial Growth Factor)
Check out our brochure on Biomarkers in Clinical Nephrology
complete ready to use ELISA kits
Literature
- Epidemiology of chronic kidney disease: an update 2022. Kidney Int Suppl (2011). Kovesdy CP. 2022 Apr;12(1):7-11. doi: 10.1016/j.kisu.2021.11.003. Epub 2022 Mar 18. PMID: 35529086; PMCID: PMC9073222.
- Chronic Kidney Disease Diagnosis and Management: A Review. Chen TK, Knicely DH, Grams ME. JAMA. 2019 Oct 1;322(13):1294-1304. doi: 10.1001/jama.2019.14745. PMID: 31573641; PMCID: PMC7015670.
- Acute kidney injury. Kellum JA, Romagnani P, Ashuntantang G, Ronco C, Zarbock A, Anders HJ. Nat Rev Dis Primers. 2021 Jul 15;7(1):52. doi: 10.1038/s41572-021-00284-z. PMID: 34267223.
- Glomerulonephritis: immunopathogenesis and immunotherapy. Anders HJ, Kitching AR, Leung N, Romagnani P.Nat Rev Immunol. 2023 Jul;23(7):453-471. doi: 10.1038/s41577-022-00816-y. Epub 2023 Jan 12. PMID: 36635359; PMCID: PMC9838307.
- Risk factors for chronic kidney disease: an update. Kazancioğlu R Kidney Int Suppl (2011). 2013 Dec;3(4):368-371. doi: 10.1038/kisup.2013.79. PMID: 25019021; PMCID: PMC4089662.
- Cardiorenal Syndrome and the Role of the Bone-Mineral Axis and Anemia. Charytan DM, Fishbane S, Malyszko J, McCullough PA, Goldsmith D Am J Kidney Dis. 2015 Aug;66(2):196-205. doi: 10.1053/j.ajkd.2014.12.016. Epub 2015 Feb 26. PMID: 25727384; PMCID: PMC4516683.
- The Effect of Diet on the Survival of Patients with Chronic Kidney Disease. Rysz J, Franczyk B, Ciałkowska-Rysz A, Gluba-Brzózka A.Nutrients. 2017 May 13;9(5):495. doi: 10.3390/nu9050495. PMID: 28505087; PMCID: PMC5452225.
Chronic kidney disease is a progressive condition that affects >10% of the general population worldwide (1). Current clinical biomarkers prove effective at advanced stages of renal impairment, limiting the timely initiation of potentially successful therapeutic interventions. There is an unmet need for more refined biomarkers capable of detecting CKD at earlier stages, thereby enhancing the prospects for patients’ outcomes.
In the last decade, the advancement in the fields of genomics, proteomics, and metabolomics have led to the identification of potential biomarker candidates that may offer important diagnostic and prognostic information in patients suffering from kidney diseases (2).
Among the current established biomarkers such as serum creatinine, albuminuria, and proteinuria, novel biomarkers for kidney diseases could potentially provide additional prognostic information. They could help to predict treatment response in various clinical settings such as acute kidney injury, transplant rejection or glomerulopathies (2).
Emerging Biomarkers in Kidney Disease
Biomedica offers a range of ELISA assay kits to reliably detect biomarkers in blood samples of patients with kidney diseases.
Endostatin, Vanin-1, Periostin, FGF23, IL-6 and more…
Complete ready-to use ELISA kits
ENDOSTATIN – a potential biomarker of renal fibrosis, chronic kidney disease (CKD), prognostic marker in acute kidney injury (AKI)
Endostatin is an extracellular matrix protein which is expressed in patients during the progression of renal fibrosis. The significant increase of serum Endostatin levels may be due to the enhanced degradation of the extracellular matrix in patients with chronic kidney disease (3, 4). Endostatin has also been studied as a prognostic marker in patients with acute kidney injury (AKI) (5) and is independently associated with incident cardiovascular events in CKD patients (6).
Endostatin can reliably be quantified in serum, plasma and urine samples:
- Endostatin ELISA | BI-20742
- Endostatin ELISA (Mouse/Rat) | BI-20742MR (7)
VANIN-1 – a potential biomarker of acute kidney injury and drug induced renal injury
Vascular non-inflammatory molecule-1 (Vanin-1) is highly expressed in the kidney (8) and has been proposed as a marker in acute kidney injury and drug induced renal injury (9). Vanin-1 has been identified as a marker of kidney damage as shown n a rat model of type 1 diabetic nephropathy (10).
Urinary Vanin-1 has been investigated in children with renal fibrosis (11) and as a predictor of acute pyelonephritis in young children with urinary tract infection (12). A recent study investigated the role of urinary Vanin-1 in kidney transplant recipients (13).
Vanin-1 can easily be measured with a conventional ELISA assay:
- Vanin-1 (urine) ELISA | BI-VAN1U
- Vanin-1 ELISA (Mouse/Rat) ELISA | BI-VAN1MR
PERIOSTIN – a potential early biomarker of renal tubular injury
Periostin is a matricellular protein that is involved in tissue remodeling and wound healing. Studies have demonstrated that the expression of Periostin in the kidney correlates with the degree of interstitial fibrosis and a decline in kidney function (15). Elevated urine Periostin levels were found in patients with type 2 diabetes which were present before the onset of microaluminuria. The authors proposed that urinary Periostin could be an early biomarker of renal tubular injury (16).
Periostin can reliably be measured in serum, plasma, and urine samples with a fully validated ELISA assay (17).
- Periostin ELISA | BI-20422
- Periostin ELISA (Mouse/Rat) | BI-20433MR
FGF23 – a potential early biomarker cardiovascular events in patients with renal-cardiovascular disease
Fibroblast growth factor 23 (FGF23) is an endocrine hormone that regulates phosphate homeostasis by modulating renal phosphate reabsorption in the kidney. Circulating FGF23 increases with declining kidney function and high FGF23 and phosphate levels are related to cardiovascular disease and mortality (18, 19).
IL-6 – a biomarker of inflammation
Interleukin-6 (IL-6) is a cytokine that plays a crucial role in inflammation and in the regulation of immune response. It is a signaling molecule that is involved in various physiological processes, including the activation of immune cells and the coordination of responses to infections or injury. IL-6 is implicated in diseases where inflammation is a prominent feature (20).
- IL-6 ELISA | BI-IL6
- HIGHLY SENSITIVE – measurable values in serum and plasma samples
- EASY – ready to use calibrators and controls
Biomedica Immunoassays
Literature
- Epidemiology of chronic kidney disease: an update 2022. Kovesdy CP. Kidney Int Suppl (2011). 2022. 12(1):7-11. doi: 10.1016/j.kisu.2021.11.003. PMID: 35529086; PMCID: PMC9073222.
- Emerging Biomarkers for Early Detection of Chronic Kidney Disease. Mizdrak M, Kumrić M, Kurir TT, Božić J. J Pers Med. 2022. 12(4):548. doi: 10.3390/jpm12040548. PMID: 35455664.
- A defective angiogenesis in chronic kidney disease. Futrakul N, Butthep P, Laohareungpanya N, Chaisuriya P, Ratanabanangkoon K. Ren Fail. 2008. 30(2):215-7. doi: 0.1080/08860220701813335. PMID: 18300124.
- Endostatin in Renal and Cardiovascular Diseases. Li M, Popovic Z, Chu C, Krämer BK, Hocher B., Kidney Dis (Basel). 2021.9;7(6):468-481. doi: 10.1159/000518221. PMID: 34901193.
- Prognostic value of dynamic plasma endostatin for the prediction of mortality in acute kidney injury: A prospective cohort study. Jia HM, Zheng Y, Han Y, Ma WL, Jiang YJ, Zheng X, Guo SY, Zhang TE, Li WX., J Int Med Res. 2020. 48(7):300060520940856. PMID: 32691651.
- Endostatin in chronic kidney disease: Associations with inflammation, vascular abnormalities, cardiovascular events and survival. Kanbay M, Afsar B, Siriopol D, Unal HU, Karaman M, Saglam M, Gezer M, Taş A, Eyileten T, Guler AK, Aydin İ, Oguz Y, Tarim K, Covic A, Yilmaz MI. Eur J Intern Med. 2016. 33:81-7. doi: 10.1016/j.ejim.2016.06.033. PMID: 27394925.
- Validation of an enzyme-linked immunosorbent assay (ELISA) for quantification of endostatin levels in mice as a biomarker of developing glomerulonephritis. Wallwitz J, Aigner P, Gadermaier E, Bauer E, Casanova E, Bauer A, Stoiber D. PLoS One. 2019. 14(8):e0220935. doi: 10.1371/journal.pone.0220935. PMID: 31404120.
- Chemical biology tools to study pantetheinases of the vanin family. Schalkwijk, J, Jansen, P. Biochem Soc Trans. 2014. 42, 1052–1055.
- Urinary Vanin-1 As a Novel Biomarker for Early Detection of Drug-Induced Acute Kidney Injury. Hosohata K et al.J Pharm Exp Ther. 2012. 341, 656–662.
- Proteomic identification of vanin-1 as a marker of kidney damage in a rat model of type 1 diabetic nephropathy. Fugmann T et al., Kidney Int. 2011. 80, 272–281.
- The Usefulness of Vanin-1 and Periostin as Markers of an Active Autoimmune Process or Renal Fibrosis in Children with IgA Nephropathy and IgA Vasculitis with Nephritis-A Pilot Study. Mizerska-Wasiak M, Płatos E, Cichoń-Kawa K, Demkow U, Pańczyk-Tomaszewska M. J Clin Med. 2022. 11(5):1265. doi: 10.3390/jcm11051265. PMID: 35268356.
- Urinary vanin-1 for predicting acute pyelonephritis in young children with urinary tract infection: a pilot study. Krzemień G, Pańczyk-Tomaszewska M, Górska E, Szmigielska A. Biomarkers. 2021. 26(4):318-324.
- Urinary vanin-1, tubular injury, and graft failure in kidney transplant recipients. Alkaff FF, Kremer D, Niekolaas TM, van den Born J, Rimbach G, Tseng TL, Berger SP, Bakker SJL, de Borst MH. Sci Rep. 2024. 4(1):2283. doi: 10.1038/s41598-024-52635-x. PMID: 38280883.
- Development of an immunoassay that reveals altered uninary Vanin-1 in human with kidney disease. Wallwitz, J., Eichinger, B., Berg, G., Gadermaier, E., Himmler, G. 2018. Nephrology Dialysis Transplantation, Volume 33, Issue suppl_1, Page i126.
- Periostin in the Kidney. Wallace DP et al., Adv Exp Med Biol. 2019, 1132:99-112.
- Periostin as a tissue and urinary biomarker of renal injury in type 2 diabetes mellitus. Satirapoj B et al., PLoS One. 2015, 17;10(4):e0124055.
- Characterization of a sandwich ELISA for the quantification of all human periostin isoforms. Gadermaier E et al., J Clin Lab Anal. 2018, 32(2):e22252.
- Higher fibroblast growth factor-23 increases the risk of all-cause and cardiovascular mortality in the community. Ärnlöv J, Carlsson AC, Sundström J, Ingelsson E, Larsson A, Lind L, Larsson TE. Kidney Int. 2013. 83(1):160-6. doi: 10.1038/ki.2012.327. PMID: 22951890.
- Fibroblast growth factor-23, cardiovascular prognosis, and benefit of angiotensin-converting enzyme inhibition in stable ischemic heart disease. Udell JA, Morrow DA, Jarolim P, Sloan S, Hoffman EB, O’Donnell TF, Vora AN, Omland T, Solomon SD, Pfeffer MA, Braunwald E, Sabatine MS. J Am Coll Cardiol. 2014. 10;63(22):2421-8, doi: 10.1016/j.jacc.2014.03.026. PMID: 24727254; PMCID: PMC4213068.
- Interleukin 6 and Cardiovascular Outcomes in Patients With Chronic Kidney Disease and Chronic Coronary Syndrome. Batra G, Ghukasyan Lakic T, Lindbäck J, Held C, White HD, Stewart RAH, Koenig W, Cannon CP, Budaj A, Hagström E, Siegbahn A, Wallentin L; STABILITY Investigators. JAMA Cardiol. 2021. 6(12):1440-1445. doi: 10.1001/jamacardio.2021.3079. PMID: 34431970; PMCID: PMC8387946.
Rare Disease Day is a global initiative held on the last day of February. It raises awareness for rare diseases to improve accessibility to medical treatment and representation for individuals diagnosed with rare diseases. It is estimated that around 300 million people worldwide are living with rare diseases.
Rare metabolic bone diseases are caused by genetic disorders that may directly or indirectly have an impact on bone structure or function (1). Other factors like hormones, tumors, diet or certain medications may also lead to abnormal growth and development of the skeleton. Some of the diseases are inherited many caused by genetic mutations. Other bone disorders are not inherited and can develop after birth.In some cases, the precise cause remains unknown.
Rare bone diseases
Rare bone diseases account for 5% of all birth defects and are an important cause of disability worldwide, yet they remain a difficult group of conditions to treat (2). It is estimated that more than 400 developmental abnormalties of the skeletal system exist (3).
The main rare metabolic bone diseases include Hypophosphatemia, Osteogenesis Imperfecta, Tumor-Induced Osteomalacia, X-Linked Hypophashatemia, and other Rare Bone Diseases (Fibrous Dysplasia, Osteopetrosis, High Bone Mass..) (4).
Biomarkers in Rare Bone Diseases
Biomarkers in Rare Bone Diseases provide a way to accelerate medical research by providing valuable insights into disease mechanisms. They play an important role in monitoring disease progression, optimizing treatments. and developing novel therapies.
NT-proCNP
C-natriuretic peptide (CNP) and its receptor, natriuretic peptide receptor-B (NPR-B) are important regulators of endochondral ossification and longitudinal bone growth (5, 6) The discovery and understanding of their physiological functions in promoting longitudinal bone growth have created opportunities for a specific targeted strategy in achondroplasia, the most common form of human dwarfism (7).
RANKL and OPG
Receptor activator of nuclear factor (NF-kappaβ) ligand (RANKL), its cellular receptor, receptor activator of NF-kappaβ (RANK), and the decoy receptor osteoprotegerin (OPG) are part of a cytokine system that regulate bone formation and resorption (8) . Denosumab is a bone anti-resorptive drug, a monoclonal antibody that binds RANKL and disrupts bone resorption. It has been approved for the treatment of osteoporosis and other bone-related disorders. The use of Denosumab in pediatric patients with Osteogenesis Imperfecta (OI), a genetic bone disorder, also known as brittle bone disease, shows decreased fractures and improved bone growth (9). A clinical trial at the National Institutes of Health found that Denosumab, significantly reduced abnormal bone turnover in adults with fibrous dysplasia, a rare disease characterized by weak, oddly shaped, or broke bones.
SCLEROSTIN
Sclerostin is a secreted protein that decreases bone formation. It binds to LRP-5 receptor on the surface of osteoblasts and consequently interferes with WNT signalling. Genetic sclerostin deficiency leads to increased bone formation and sclerotic bone disorders. Sclerostin inhibition is being evaluated as a potential approach to increase bone mass in Osteogenesis Imperfecta (10).
FGF23
Fibroblast growth factor 23 (FGF23) is a hormone that is produced by bone. It regulates serum phosphate levels by suppressing phosphate reabsorption in the kidney. Excessive actions of FGF23 are responsible for different kinds of hypophosphatemic rickets as found in X-linked hypophosphatemia (XLH) and osteomalacia. XLH is characterized by deformities of the lower limb and short stature. An anti-fibroblast growth factor-23 (FGF23) monoclonal antibody (Burosumab) has been approved as a novel treatment for hypophopshatemic rickets (11).
Measurement of FGF23 is a critical tool to assist in the evaluation and diagnosis of hypophosphatemic conditions (12, 13).
The second most common genetic form of hypophosphatemic rickets after XLH, is autosomal-dominant hypophosphatemic rickets (ADHR). ADHR is caused by specific mutations in the FGF23 gene.
FGF23 can reliably be measured with an immunoassay (14).
FGF23 ELISAs (c-term FGF23, cat. no BI-20702) (intact FGF23, cat. no. BI-20700)
- MULTI-USE for serum and plasma samples
- TRUSTED in over 60 citations
FGF23 – Test
Biomarker ELISA assay kits from BIOMEDICA
Complete ready-to use ELISA kits
NT-proCNP ELISA (cat. no. BI-20812)
- CONVENIENT – low sample volume – 20 µl / well
- RELIABLE – validated following international quality guidelines
- TRUSTED – widely cited in over 40 publications (click here for full list)
- MULTI-USE – for human serum and plasma samples; protocols for non-human samples (e.g. rat).
Citations Selection of NT-proCNP ELISA citations related to skeletal disorders:
- Plasma C-Type Natriuretic Peptide: Emerging Applications in Disorders of Skeletal Growth. Espiner E et al.,Horm Res Paediatr. 2018. 90(6):345-357. doi: 10.1159/000496544. PMID: 30844819.
- Rats deficient C-type natriuretic peptide suffer from impaired skeletal growth without early death. Fujii T et al., PLoS One. 2018. 22;13(3):e0194812. doi: 10.1371/journal.pone.0194812. PMID: 29566041.
- Serum NT-proCNP levels increased after initiation of GH treatment in patients with achondroplasia/hypochondroplasia. Kubota T etal., Clin Endocrinol (Oxf). 2016 Jun;84(6):845-50. doi: 10.1111/cen.13025. Epub 2016 Feb 25. PMID: 26814021.
- Acromesomelic dysplasia, type maroteaux caused by novel loss-of-function mutations of the NPR2 gene: Three case reports. Wang W et al., 2016. Am J Med Genet A 170A, 426–434.
- Skeletal overgrowth syndrome caused by overexpression of C-type natriuretic peptide in a girl with balanced chromosomal translocation, t(1;2)(q41;q37.1). Ko J et al., 2015. Am J Med Genet A 167A, 1033–1038.
RANKL ELISA (cat. no. BI-20462)
- Highly sensitive – measurable concentrations in healthy subjects
- Only assay that measures free, uncomplexed, soluble RANK Ligand
- TRUSTED in over 300 citations
- CONVENIENT- ready to use liquid calibrators and controls
- EFFICIENT – low sample volume 20µl / well
- TRUSTED in over 250 citations
Literature
- Genetic approaches to metabolic bone diseases. Hannan FM, Newey PJ, Whyte MP, Thakker RV. Br J Clin Pharmacol. 2019. 85(6):1147-1160. doi: 10.1111/bcp.13803. PMID: 30357886; PMCID: PMC6533455.
- The evolving therapeutic landscape of genetic skeletal disorders. Sabir, A.H., Cole, T. Orphanet J Rare Dis 14, 300 (2019).
- Changes in skeletal dysplasia nosology. Jurcă MC, Jurcă SI, Mirodot F, Bercea B, Severin EM, Bembea M, Jurcă AD. Rom J Morphol Embryol. 2021. 2(3):689-696. doi: 10.47162/RJME.62.3.05. PMID: 35263396; PMCID: PMC9019670.
- Atlas of rare genetic metabolic bone diseases. IOF, 2024.
- Natriuretic peptide regulation of endochondral ossification. Evidence for possible roles of the C-type natriuretic peptide/guanylyl cyclase-B pathway. Yasoda A, Ogawa Y, Suda M, Tamura N, Mori K, Sakuma Y, Chusho H, Shiota K, Tanaka K, Nakao K J Biol Chem.1998. 8;273(19):11695-700. doi: 10.1074/jbc.273.19.11695. PMID: 9565590.
- Cyclic GMP-dependent protein kinase II plays a critical role in C-type natriuretic peptide-mediated endochondral ossification. Miyazawa T, Ogawa Y, Chusho H, Yasoda A, Tamura N, Komatsu Y, Pfeifer A, Hofmann F, Nakao K. Endocrinology. 2002. 143(9):3604-10. doi: 10.1210/en.2002-220307. PMID: 12193576.
- Optimal management of complications associated with achondroplasia. Ireland PJ, Pacey V, Zankl A, Edwards P, Johnston LM, Savarirayan R Appl Clin Genet. 2014. 7:117-25. doi: 10.2147/TACG.S51485. PMID: 25053890; PMCID: PMC4104450.
- C-type natriuretic peptide regulates endochondral bone growth through p38 MAP kinase-dependent and -independent pathways. Agoston H, Khan S, James CG, Gillespie JR, Serra R, Stanton LA, Beier F BMC Dev Biol. 2007. 7:18. doi: 10.1186/1471-213X-7-18. PMID: 17374144; PMCID: PMC1847438.
- Role of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in bone cell biology. Hofbauer LC, Heufelder AE. J Mol Med (Berl). 2001 Jun;79(5-6):243-53. doi: 10.1007/s001090100226. PMID: 11485016.
- Effect of denosumab on the growing skeleton in osteogenesis imperfecta. Hoyer-Kuhn H, Semler O, Schoenau E.J Clin Endocrinol Metab. 2014. 99(11):3954-5. doi: 10.1210/jc.2014-3072. PMID: 25148238.
- Effect of sclerostin inactivation in a mouse model of severe dominant osteogenesis imperfecta. Marulanda J, Tauer JT, Boraschi-Diaz I, Bardai G, Rauch F. Sci Rep. 2023. 13(1):5010. doi: 10.1038/s41598-023-32221-3. PMID: 36973504; PMCID: PMC10043013.
- Hereditary Metabolic Bone Diseases: A Review of Pathogenesis, Diagnosis and Management. Charoenngam N, Nasr A, Shirvani A, Holick MF.Genes (Basel). 2022. 13(10):1880. doi: 10.3390/genes13101880. PMID: 36292765; PMCID: PMC9601711.
- Determination of FGF23 Levels for the Diagnosis of FGF23-Mediated Hypophosphatemia. Hartley IR, Gafni RI, Roszko KL, Brown SM, de Castro LF, Saikali A, Ferreira CR, Gahl WA, Pacak K, Blau JE, Boyce AM, Salusky IB, Collins MT, Florenzano P. J Bone Miner Res. 2022. 37(11):2174-2185. doi: 10.1002/jbmr.4702. PMID: 36093861; PMCID: PMC9712269.
- The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Heijboer AC, Cavalier E Calcif Tissue Int. 2023. 112(2):258-270. doi: 10.1007/s00223-022-00987-9. PMID: 35665817; PMCID: PMC9859838.
Heart failure (HF) is a leading cause of death in individuals with chronic kidney disease (CKD). CKD is also known as a “silent killer” as many people live with the disease without developing symptoms. CKD is a progressive disease and it is estimated that around 10% of the adult general population worldwide is affected (1). A large number of individuals are undiagnosed as demonstrated in a study from the UK in 3200 participants aged over 60 years (2).
FGF23 and Risk of Heart Failure
What is CKD?
Chronic kidney disease is a condition where kidney function gradually declines. The kidneys play a crucial role in filtering waste from the blood with the help of nephrons. A constant increase in damaged and non-functioning nephrons lead to the progression of the disease.
How does CKD cause Heart Failure?
Impaired kidney function places an additional strain on the heart, as it needs to pump harder to get blood to the kidneys. Changes in blood pressure is also a complication in CKD that can ultimately lead to heart disease. One suggested mechanism for the development of HF in CKD patients is linked to highly elevated levels of FGF23 (3).
What is the role of FGF23 in Heart Failure?
Fibroblast growth factor 23 (FGF 23) is a bone-derived hormone that regulates phosphate and vitamin D. In individuals with CKD, circulating FGF23 blood levels gradually increase with declining kidney function. In patients with end-stage renal disease, FGF23 levels are highly elevated up to 1000-fold above the normal range (4).
Numerous studies indicate that high FGF23 concentrations have a direct effect on the heart and the cardiovascular system. Elevated FGF23 levels have been linked to an increased risk of cardiovascular events, including heart failure, cardiovascular mortality, and left ventricular hypertrophy (5, 6). The precise mechanisms by which FGF23 impacts the heart are still being explored. One potential pathway could be linked to the discovery of FGF23 receptors expressed in the heart by cardiomyocytes. Specific binding of FGF23 to these receptors may directly induce hypertrophy. Additional pathways are under investigation.
In a recent large multicenter prospective cohort study in over 3500 participants, researchers investigated the relationship between elevated FGF23 levels and the risk of HF in individuals with varies HF subtypes. The authors concluded that elevated FGF23 levels were associated with increased risks for all HF subtypes highlighting the need for further research on FGF23 as a potential target in HF prevention in individuals with chronic kidney disease (3).
What are the methods for measuring FGF23?
Circulating blood FGF23 levels can be measured with immunoassays. A commonly employed technique is the ELISA assay (enzyme linked immunosorbent assay).
FGF23 ELISA assays employ specific antibodies designed to detect FGF23 present in the sample (serum / plasma). Currently, two commercially available assays are used for the measurement of circulating FGF23 in blood samples (7):
- ELISA for measuring Intact FGF23
The full length, biologically active form of the hormone is intact FGF23 (iFGF23), consisting of the complete and full-length FGF23 protein structure without enzymatical cleavage. The ELISA utilizes two antibodies that target the N-terminal part and the C-terminal domain of the FGF23 molecule, respectively.
FGF23 intact ELISA assay – binding region of antibodies utilized in the BIOMEDICA kit
- ELISA for measuring FGF23 C-terminal
C-terminal fragments of FGF23 (cFGF23) result from the enzymatic cleavage of the intact FGF23 molecule. The ELISA utilizes two antibodies that bind to epitopes that are located in the c-terminal domain of the FGF23 molecule.
FGF23 (C-terminal) ELISA assay – binding region of antibodies utilized in the BIOMEDICA kit
OF NOTE: all FGF23 c-terminal assays that are commercially available detect both c-terminal FGF23 fragments as well as the intact FGF23 molecule (7).
BIOMEDICA offers two ELISA kits that reliably quantify FGF23 concentrations in human serum and plasma samples:
FGF23 – complete ready to use ELISA kits – User-Friendly and Reliable
- FGF23 intact ELISA (cat. no. BI-20700)
- FGF23 (C-terminal) ELISA (cat. no. BI-20702)
BIOMEDICA FGF23 intact ELISA assay (#BI-20700)
BIOMEDICA FGF23 multimatrix (C-terminal) ELISA assay (#BI-20702)
- Validated FGF23 ELISA kits according to international quality guidelines
- Numerous citations /references in top-ranking journals
FGF23 ELISA assays are Developed & Manufactured by Biomedica
Literature
- Epidemiology of chronic kidney disease: an update 2022. Kovesdy CP. Kidney Int Suppl (2011). 2022. 12(1):7-11. PMID: 35529086; PMCID: PMC9073222
- Prevalence of chronic kidney disease in the community using data from OxRen: a UK population-based cohort study. Hirst JAet al., Br J Gen Pract. 2020. 26;70(693):e285-e293. PMID: 32041766; PMCID: PMC7015167.
- Chronic Renal Insufficiency Cohort (CRIC) study investigators. Fibroblast Growth Factor 23 and Risk of Heart Failure Subtype: The CRIC (Chronic Renal Insufficiency Cohort) Study. Leidner Aset al.,Kidney Med. 2023. 5(11):100723. PMID: 37915961; PMCID: PMC10616385.
- Fibroblast growth factor-23 mitigates hyperphosphatemia but accentuates calcitriol deficiency in chronic kidney disease. Gutierrez O et al., Journal of the American Society of Nephrology. 2005;16(7):2205–2215.
- FGF23 and Cardiovascular Risk. Prié D. Ann Endocrinol (Paris). 2021. 82(3-4):141-143. PMID: 32950228.
- Paracrine Effects of FGF23 on the Heart. Front Endocrinol (Lausanne). Leifheit-Nestler M, Haffner D 2018. 28;9:278. PMID: 29892269; PMCID: PMC5985311.
- The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Calcif Tissue Int. Heijboer AC, Cavalier E. 2023 Feb;112(2):258-270. PMID: 35665817; PMCID: PMC9859838.
The enzyme-linked immunosorbent assay (ELISA or EIA) is a laboratory method to detect and quantify the presence of a protein in biological samples (1, 2).
ELISA Assay Principle
When selecting an ELISA kit, researchers are often confronted with the question which assay to choose of the many commercially available kits.
It can be a challenge! Here are a few hints that may help.
HOW TO CHOOSE THE RIGHT ELISA KIT
As a general rule, before purchasing an assay, always read the protocol booklet (instructions for use – IFU or package insert) in detail. This should ensure that the kit will be suitable for your requirements. Check out the following:
1. ANALYTE
Which protein biomarker will you be measuring? Be sure to use the correct term during your search. Some biomarkers have alternative names (e.g. Sclerostin or SOST ELISA (SOST is actually the name for the gene that encodes Sclerostin).
2. SPECIES – SPECIFICITY – CROSS REACTIVITY
Verify if the assay can be used in the respective model such as e.g. human, rat, or mouse. Due to high homology between species, some ELISA kits work both in humans and in different animal species. As an example the biomarker ELISA kit for NT-proANP was developed for human use but due to the high sequence homology between species, the kit is successfully used to measure NT-proANP as a cardiac safety biomarker in various animal models (rat, mouse, rabbit, monkey).
3. SAMPLE TYPE
What is the sample type (matrix) you´ll be using (e.g. serum, EDTA-plasma, heparin-plasma, citrate-plasma, cell culture supernatants, urine..) ?
Verify if the assay is compatible for your sample type: check the package insert and, if available, check if there are validation data showing results (often found on the manufacturers website).
Of note: analysis of some biomarkers in the “wrong” matrix can lead to “false” results due to a matrix effect.
4. SAMPLE VOLUME
Check the amount of sample required per well (calculate volume to measure your samples in duplicates). Low sample volumes and precious samples are often a selection criterium.
5. SENSITIVTY – BIOMARKER CONCENTRATIONS TO BE EXPECTED
Before choosing an assay, look into the validation data of the kit (often documented in the ELISA protocol booklet or in the validation data files).
Reference values and pathological values in serum and/or plasma of the biomarker of interest are sometimes documented as well. These data can be helpful in selecting an appropriate assay. In some cases samples may require a pre-dilution. Therefore, always verify if the dilution buffer (assay buffer) is included in the kit or ask the assay developer for their input.
Of note: assays offering high sensitivity offer a different dynamic range than assays with a lower sensitivity. The dynamic range of an assay indicates the range of concentrations over which an assay can accurately quantify the analyte.
6. ASSAY PERFORMANCE – ASSAY VALIDATION
Careful evaluation of the assay´s performance characteristics is important in selecting an ELISA kit.
Choose an assay that has gone through a rigorous validation process. Check out if you can find data on the following performance characteristics:
- Accuracy- detection of a protein biomarker in clinical samples.
- Dilution linearity and parallelism – recovery of the analyte of interest in diluted samples
- Specificity & cross-reactivity – making sure that you detect only the analyte of interest
- Precision – within-run and in-between run precision – ensuring precise and reproducible results within an across assay lots
- Calibration – ensures consistent performance over the range of the assay of the calibration curve
- Sample stability – ensures the stability of the analyte of interest (e.g. exposure of real samples to multiple freeze-thaw cycles, stability at room temperature..).
ELISA Assay – microtiter plate
7. KIT COMPONENTS
Verify if the contents of the ELISA kit includes all the necessary components e.g. controls, assay dilution buffer. Consider storage requirements such as temperature sensitivity and expiration date.
8. CITATIONS & REFERENCES
Check if there are citations on the manufacturers website for the specific ELISA kit. Look into publications and seek feedback from researchers who have used the assay you are considering.
9. PRODUCT ORIGIN
Verify if the kit supplier is the kit manufacturer. More and more kits are repacked and are sold under different names, although it is always the same kit.
ELISA kit manufacturers will more likely give you qualified support as they “know” their product (e.g. availability of additional calibrators, controls, buffers.., technical know-how on the kit..).
10. CUSTOMER SUPPORT
Verify if the kit provider can provide timely and helpful customer service.
Related Literature
1.Enzyme-Linked Immunosorbent Assay: Types and Applications. Hayrapetyan H, Tran T, Tellez-Corrales E, Madiraju C. Methods Mol Biol. 2023;2612:1-17. doi: 10.1007/978-1-0716-2903-1_1. PMID: 36795355.
2. Enzyme Immunoassay (EIA)/Enzyme-Linked Immunosorbent Assay (ELISA). Lequin RM, Clinical Chemistry. 2005; 51, 12: 2415-2418.
February is marked as “Heart Month” drawing attention to heart disease and its prevention. Cardiovascular diseases (CVD) are the leading cause of death globally with an estimated toll of 17.9 million people per year (1).
Unhealthy diet, physical inactivity, tobacco use are only some of the behavioral risk factors associated with heart disease. Health conditions that increase the risk include obesity, elevated blood sugar levels, high blood pressure, high low-density lipoprotein (LDL) cholesterol, and oxidative stress.
Biomarkers of Oxidative Stress in Heart Disease
What is oxidative stress?
Oxiidative Stress- balance antioxidants and free radicals
Oxidative stress is caused by an imbalance between the production of free radicals (reactive oxygen species- ROS) and antioxidant defenses (endogenous antioxidant capacity). ROSs are generated as metabolic by-products in cells that can be stimulated by a variety of agents including pollutants, heavy metals, tobacco, drugs and many other. It has been postulated that oxidative stress can be responsible in the onset and progression of several diseases (e.g. cancer, diabetes, metabolic disorders, atherosclerosis, and cardiovascular diseases) (2).
Oxidized LDL (oxLDL) and Anti-oxidized LDL Autoantibodies
Accumulation of oxidized-low density lipoproteins (oxLDL) in the blood vessels triggers the onset of atherosclerosis which is a key element of the development of cardiovascular disease (CVD). The term atherosclerosis is of Greek origin, meaning thickening of the intimal layer of arteries and accumulation of fat (3). The process of atherosclerosis is initiated by LDL particles that lie in the sub-endothelial space of arteries (4). Thus, atherosclerosis is a disease of the vascular intima as a result of lipid oxidation. In the later stages of disease progression, plaques build up that rupture resulting in thrombosis (blockage of blood flow). oxLDL induces inflammatory responses in macrophages and dendritic cells (5).
Autoantibodies against oxidatively modified LDL (anti-oxLDL Ab) have been detected in patients with atherosclerosis as well as in healthy individuals (6). It has been suggested that they mirror the occurrence of oxidation processes taking place in vivo. Anti-oxLDL Ab can be measured in human blood samples with a conventional ELISA assay (6-8). Numerous studies have implied that autoantibodies to oxLDL may allow improved CV risk stratification (6).
How can oxidative status / oxidative stress be measured?
Oxidative stress can be measured indirectly by different techniques.
Example of a BIOMEDICA ELISA kit
1.MEASUREMENT OF ANTI-OXIDIZED LDL AUTOANTIBODIES
ELISA for the detection of Anti-oxidized LDL Autoantibodies (oLAB ) | BI-20032
- Sample type: serum
- Sample volume 50µl/well
- Incubation time: 1.5 h / 30 min / 15 min
- Detection range: 0 – 1200 mU / ml
- Sensitivity: 48 mU / ml
- Precision: In-between-run (n=5): ≤ 8 % CV, Within-run (n=8): ≤ 4 % CV
- Use: Research use only
- Widely cited in over 70 publications
2. MEASUREMENT OF BIOLOGICAL PEROXIDES
Oxidative Stress Test – OXYSTAT Assay | BI-5007
Quick and easy assay to measure total peroxides in biological fluids.
- Method: colorimetric assay, 96 wells
- Sample type: serum, plasma, biological fluids
- Sample volume 10µl/well
- Assay time: 15 min
- Detection range: 0 – 660 µmol/l
- Sensitivity: 7 µmol/l
- Use: Research use only
- Widely cited in over 50 publications
Results show a direct correlation between free radicals and circulating biological peroxides and thus allow the characterization of the oxidative status in biological samples.
Literature
- WHO-international health topics-cardiovascular diseases.
- Reactive oxygen species in the vasculature: molecular and cellular mechanisms. Taniyama Y, Griendling KK. Hypertension. 2003 Dec;42(6):1075-81. doi: 10.1161/01.HYP.0000100443.09293.4F. Epub 2003 Oct 27. PMID: 14581295.
- Atherosclerosis: process, indicators, risk factors and new hopes. Rafieian-Kopaei M, Setorki M, Doudi M, Baradaran A, Nasri H. Int J Prev Med. 2014 Aug;5(8):927-46. PMID: 25489440; PMCID: PMC4258672.
- Oxidized LDL and anti-oxidized LDL antibodies in atherosclerosis – Novel insights and future directions in diagnosis and therapy. Hartley A, Haskard D, Khamis R. Trends Cardiovasc Med. 2019 Jan;29(1):22-26. doi: 10.1016/j.tcm.2018.05.010. Epub 2018 Jun 4. PMID: 29934015.
- How Oxidized Low-Density Lipoprotein Activates Inflammatory Responses. Rhoads JP, Major AS. Crit Rev Immunol. 2018;38(4):333-342. doi: 10.1615/CritRevImmunol.2018026483. PMID: 30806246; PMCID: PMC6527110.
- Anti-Oxidized LDL Antibodies and Coronary Artery Disease: A Systematic Review. van den Berg VJ, Vroegindewey MM, Kardys I, Boersma E, Haskard D, Hartley A, Khamis R. Antioxidants (Basel). 2019 Oct 15;8(10):484. doi: 10.3390/antiox8100484. PMID: 31618991; PMCID: PMC6826549
- Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou E, Vourli G, Nounopoulos C, Zachari A, Dionyssiou-Asteriou A. Free Radic Res. 2005 Apr;39(4):419-29. doi: 10.1080/10715760500072156. PMID: 16028367.
- Antibodies to oxidized low-density lipoprotein in patients following coronary artery revascularization. Miller ER 3rd, Erlinger TP, Blumenthal RS, Margolis S, Allen JK. Coron Artery Dis. 2003 Apr;14(2):163-9. doi: 10.1097/00019501-200304000-00009. PMID: 12655280.
Biomedica´s Osteoprotegerin (OPG) and Fibroblast Growth Factor (FGF23) assays were highlighted in the German Chronic Kidney Disease (CKD) cohort study to investigate the prognostic value of a total of nine mineral and bone disease biomarkers (MBD) (1). Brief, 4246 participants were enrolled and baseline serum samples of the MBD markers were prospectively observed for a median time of 6.5 years. The markers were compared to study the association of each biomarker with adverse cardiovascular (CV) outcomes and mortality. Of the nine biomarkers examined, Osteoprotegerin (OPG) best identified CKD patients who had the highest risk. The findings mirror the link between bone metabolism and CV disease in CKD patients and may help clinicians to identify patients who are at high risk for CV disease (1).
OPG a marker of cardiovascular mortality in CKD
The Kidney, Bone, and Heart Connection
Chronic Kidney Disease (CKD), is a condition involving a gradual loss of kidney function. The kidneys lose their ability to filter toxins and extra fluid from the blood. There is no cure for this progressive disease and it is associated with a high morbidity and mortality rate that occurs especially in people with diabetes and hypertension (2). Worldwide, it is estimated that CKD is present in one out of ten adults (3).
Bone Disorders in Chronic Kidney Disease
Other important roles of the kidney include maintaining bone health and balancing important minerals in the body. A majority of patients with CKD have an elevated risk of developing disturbances of bone and mineral metabolism that lead to bone lesions (4). Thus, mineral an bone disorders (MBD) with biochemical and hormonal alterations are part of the complications associated with the progression of CKD (5).
Cardiovascular Events in Chronic Kidney Disease (CKD)
Mineral and bone disorder in CKD can not only affect the bones but also other organs including the heart and blood vessels. It is well recognized that both atherosclerosis and arteriosclerosis are accelerated in patients with kidney failure (6). The high cardiovascular morbidity and mortality in CKD patients is mainly caused by progressive vascular calcification.
Osteoprotegerin (OPG) a Cardiovascular Risk Factor in CKD
Osteoprotegerin (OPG) is a protein that regulates bone mass by inhibiting bone resorption. Apart from its traditional role in bone, OPG plays an important role in vascular disease and calcification as it interacts with endothelial and smooth muscle cells (7).
In a large prospective, population based study, severity, initiation, and progression of atherosclerosis was assessed in carotid arteries (8). The researchers demonstrated that OPG was an independent risk factor for the progression of atherosclerosis and onset of cardiovascular disease (CV) (8).
Serum OPG has also been shown to be a predictor of progression of atherosclerosis and coronary calcification in hemodialysis patients (9) and OPG serum levels increase with CKD disease progression and are associated with mortality in these patients (10). Moreover, high OPG levels are also associated with CV events in the general population as demonstrated in a large community based cohort (10). These data strongly reinforce OPG as marker for CV disease risk factor burden and predictor of CVD and mortality in the community (11).
Finally, the data from the recent German Chronic Kidney Disease cohort study investigating the prognostic value of nine mineral and bone disease associated biomarkers (MBD) showed that only serum OPG levels consistently identified the CKD patients who had the highest risk for adverse CV outcomes and mortality (1).
Osteoprotegerin (OPG) and Fibroblast Growth Factor (FGF23) can reliably be measured with a conventional ELISA assay
Biomedica OPG ELISA (cat. no. BI-20403)
- Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
- Sample: Serum, plasma (EDTA, citrate, heparin)
- Sample volume: 20μl / well
- Detection range: 1.25-20 pmol/L (= 25 – 400 pg/mL)
- Sensitivity: 0.07 pmol/L (= 1.4 pg/mL)
- Incubation time: 4 h + 1 h + 30 min (room temperature)
- Precision: In-between-run (n=12): ≤ 5 % CV: Within-run (n=5): ≤ 3 % CV
- Widely cited in over 250 publications!
Biomedica FGF23 (C-terminal) ELISA (cat. no. BI-20702)
- Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
- Sample: Serum, plasma (EDTA, citrate, heparin)
- Sample volume: 50μl / well
- Detection range: 0.2 – 20 pmol/mL (= 1.54 – 150.4 pg/mL)
- Sensitivity: 0.08 pmol/L (= 0.6 pg/mL)
- Incubation time: 20-24 h + 1 h+ 30 min (room temperature)
- Precision: In-between (n=10): ≤ 10 % CV; Within-run (n=6): ≤ 12 % CV
Publications
- Association of mineral and bone biomarkers with adverse cardiovascular outcomes and mortality in the German Chronic Kidney Disease (GCKD) cohort. Reimer KC, Nadal J, Meiselbach H, Schmid M, Schultheiss UT, Kotsis F, Stockmann H, Friedrich N, Nauck M, Krane V, Eckardt KU, Schneider MP, Kramann R, Floege J, Saritas T; GCKD study investigators. Bone Res. 2023 Oct 20;11(1):52. doi: 10.1038/s41413-023-00291-8. PMID: 37857629; PMCID: PMC10587182.
- Chronic kidney disease. Kalantar-Zadeh K, Jafar TH, Nitsch D, Neuen BL, Perkovic V. Lancet. 2021 Aug 28;398(10302):786-802. doi: 10.1016/S0140-6736(21)00519-5. Epub 2021 Jun 24. PMID: 34175022.
- Prevalence, outcomes, and cost of chronic kidney disease in a contemporary population of 2·4 million patients from 11 countries: The CaReMe CKD study. Sundström J, Bodegard J, Bollmann A, Vervloet MG, Mark PB, Karasik A, Taveira-Gomes T, Botana M, Birkeland KI, Thuresson M, Jäger L, Sood MM, VanPottelbergh G, Tangri N; CaReMe CKD Investigators. Lancet Reg Health Eur. 2022. 20:100438. doi: 10.1016/j.lanepe.2022.100438. PMID: 36090671; PMCID: PMC9459126.
- Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD): Current Perspectives. Waziri B, Duarte R, Naicker S. Int J Nephrol Renovasc Dis. 2019 Dec 24;12:263-276. doi: 10.2147/IJNRD.S191156. PMID: 31920363; PMCID: PMC6935280.
- Mineral Bone Disorders in Kidney Disease Patients: The Ever-Current Topic. Hu L, Napoletano A, Provenzano M, Garofalo C, Bini C, Comai G, La Manna G. Int J Mol Sci. 2022 Oct 13;23(20):12223. doi: 10.3390/ijms232012223. PMID: 36293076; PMCID: PMC9603742
- Role of Chronic Kidney Disease (CKD)-Mineral and Bone Disorder (MBD) in the Pathogenesis of Cardiovascular Disease in CKD. Yamada S, Nakano T J Atheroscler Thromb. 2023 Aug 1;30(8):835-850. doi: 10.5551/jat.RV22006. Epub 2023 May 30. PMID: 37258233; PMCID: PMC10406631.
- Role of crosstalk between endothelial cells and smooth muscle cells in vascular calcification in chronic kidney disease. Zhang YX, Tang RN, Wang LT, Liu BC. Cell Prolif. 2021. 54(3):e12980. doi: 10.1111/cpr.12980. Epub 2021 Jan 27. PMID: 33502070; PMCID: PMC7941222.
- Osteoprotegerin is a risk factor for progressive atherosclerosis and cardiovascular disease. Kiechl S, Schett G, Wenning G, Redlich K, Oberhollenzer M, Mayr A, Santer P, Smolen J, Poewe W, Willeit J. Circulation. .109(18):2175-80. doi: 10.1161/01.CIR.0000127957.43874.BB. Epub 2004 Apr 26. PMID: 15117849.
- Serum osteoprotegerin is a predictor of progression of atherosclerosis and coronary calcification in hemodialysis patients. Kurnatowska I, Grzelak P, Kaczmarska M, Stefańczyk L, Nowicki M. Nephron Clin Pract. 2011;117(4):c297-304. doi: 10.1159/000321169. Epub 2010 Sep 22. PMID: 20861651.
- Circulating Osteoprotegerin in Chronic Kidney Disease and All-Cause Mortality. Kamińska J, Stopiński M, Mucha K, Pac M, Gołębiowski M, Niewczas MA, Pączek L, Foroncewicz B. Int J Gen Med. 2021 Jun 9;14:2413-2420. doi: 10.2147/IJGM.S302251. PMID: 34135625; PMCID: PMC8200134.
- Biomarkers of the osteoprotegerin pathway: clinical correlates, subclinical disease, incident cardiovascular disease, and mortality. Lieb W, Gona P, Larson MG, Massaro JM, Lipinska I, Keaney JF Jr, Rong J, Corey D, Hoffmann U, Fox CS, Vasan RS, Benjamin EJ, O’Donnell CJ, Kathiresan S. Arterioscler Thromb Vasc Biol. 2010 Sep;30(9):1849-54. doi: 10.1161/ATVBAHA.109.199661. Epub 2010 May 6. PMID: 20448212; PMCID: PMC3039214.
- Association of mineral and bone biomarkers with adverse cardiovascular outcomes and mortality in the German Chronic Kidney Disease (GCKD) cohort. Reimer KC, Nadal J, Meiselbach H, Schmid M, Schultheiss UT, Kotsis F, Stockmann H, Friedrich N, Nauck M, Krane V, Eckardt KU, Schneider MP, Kramann R, Floege J, Saritas T; GCKD study investigators. Bone Res. 2023 Oct 20;11(1):52. doi: 10.1038/s41413-023-00291-8. PMID: 37857629; PMCID: PMC10587182.