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• FGF23 and Sclerostin - novel biomarkers in diabetic kidney disease

November is “Diabetes Awareness Month” raising attention to this fast growing and life-threating epidemic. Patients suffering from diabetes have a risk of additional health complications, including heart disease, strokes, and diabetic kidney disease (DKD). People who develop DKD mostly have few symptoms in the early stage of the disease, although the risk of developing severe kidney damage is very high. High blood sugar levels may damage the small blood vessels in the kidney leading to kidney damage, kidney failure, and high blood pressure (1). 

FGF23 and Sclerostin – novel biomarkers in diabetic kidney disease

Traditionally, the bone is regarded as a structural organ that gives the human body support and facilitates physical movement. However, bone is also a source of various hormones including fibroblast growth factor 23 and sclerostin that play an important role in regulating glucose metabolism and DKD (2).

FGF23 and Sclerostin  – novel biomarkers in diabetic kidney disease

 

FGF23 and Sclerostin – bone derived hormones regulate glucose metabolism

 

Fibroblast growth factor 23 (FGF23) is a bone-derived protein that regulates phosphate metabolism, by inhibiting renal phosphate reabsorption. There is increasing evidence that FGF23 plays a role in type 2 diabetes (T2DM), as FGF23 levels are elevated in these patients, even in individuals with preserved kidney function when compared to the general population (3). Phosphate independent effects on FGF23 following glucose loading were shown in a recent study demonstrating that FGF23 is associated with glucose, insulin and proinsulin levels, as well as obesity (4 ). Furthermore, FGF23 has also been shown to be associated with the development of gestational diabetes mellitus (5).

Sclerostin is a protein that is produced by bone cells that inhibits bone formation. Recent research suggests that Sclerostin also plays a role in lipid and glucose metabolism as serum sclerostin is negatively associated with insulin sensitivity as measured in obese, but not lean women (5). Sclerostin levels have also been shown to be increased in individuals with prediabetes (6).

FGF23 and Sclerostin can reliable by measured with conventional ELISA assays from BIOMEDICA.

FGF23 (C-terminal), #BI-20700 and FGF23 intact ELISA, #BI-20700

• RELIABLE – validated following international quality guidelines
• CITED in over 60 publications
• EASY – 8 standards and 2 controls included
• For SERUM & PLASMA samples
• HIGH QUALITY guaranteed!

 

Sclerostin ELISA, #BI-20492

• MOST referenced in more than 280 citations
• LOW sample volume – 20µl sample /well
• For SERUM & PLASMA samples
• RELIABLE – full validation package

 

Bioactive Sclerostin ELISA, BI-20472

• CHARACTERIZED ANTIBODIES – targeting the receptor binding region
• RIGOROUSLY validated for clinical samples
• LOW sample volume – 20µl sample /well

 

Literature

1. Renal microvascular disease predicts renal function in diabetes. Futrakul N, Futrakul P. Ren Fail. 2012;34(1):126-9. doi: 10.3109/0886022X.2011.623490. Epub 2011 Oct 20. PMID: 22010784.
2. The Emerging Role of Bone-Derived Hormones in Diabetes Mellitus and Diabetic Kidney Disease. Li Y, Gu Z, Wang J, Wang Y, Chen X, Dong B. Front Endocrinol (Lausanne). 2022 Jul 11;13:938830. doi: 10.3389/fendo.2022.938830. PMID: 35966090; PMCID: PMC9367194.
3. Earlier onset and greater severity of disordered mineral metabolism in diabetic patients with chronic kidney disease Wahl P., Xie H., Scialla J., Anderson C.A.M., Bellovich K., Brecklin C.et al.. (2012)  Diabetes Care 35, 994–1001 10.2337/dc11-2235 
4. Fibroblast Growth Factor 23, Glucose Homeostasis, and Incident Diabetes: Findings of 2 Cohort Studies, Amarens van der Vaart, Coby Eelderink, André P van Beek, Stephan J L Bakker, Peter R van Dijk, Martin H de Borst, The Journal of Clinical Endocrinology & Metabolism, Volume 108, Issue 10, October 2023, Pages e971–e978, https://doi.org/10.1210/clinem/dgad246.
5. Fibroblast growth factor 23 is associated with the development of gestational diabetes mellitus. Hocher CF, Chen X, Zuo J, Horvathova K, Hocher B, Krämer BK, Chu C. Diabetes Metab Res Rev. 2023 Aug 8:e3704. doi: 10.1002/dmrr.3704. Epub ahead of print. PMID: 37553983.
6. Serum sclerostin is negatively associated with insulin sensitivity in obese but not lean women. Aznou A, Meijer R, van Raalte D, den Heijer M, Heijboer A, de Jongh R. Endocr Connect. 2021 Feb;10(2):131-138. doi: 10.1530/EC-20-0535. PMID: 33480863; PMCID: PMC7983521.
7. Sclerostin and Insulin Resistance in Prediabetes: Evidence of a Cross Talk Between Bone and Glucose Metabolism. Daniele G, Winnier D, Mari A, Bruder J, Fourcaudot M, Pengou Z, Tripathy D, Jenkinson C, Folli F. Diabetes Care. 2015 Aug;38(8):1509-17. doi: 10.2337/dc14-2989. Epub 2015 Jun 17. PMID: 26084344.

Cigarette smoking is involved in the development of cardiovascular disease (CVD). It leads to changes in a number of biomarkers reflecting inflammation, oxidative stress, endothelial function, and lipid levels (1, 2) . A recent study investigated changes in biomarkers in individuals who stopped smoking (1). One of the biomarkers to measure oxidative stress in serum of individuals who quit smoking was “anti-oxidized low-density lipoprotein (anti-oxLDL) antibodies”. The anti-oxLDL antibodies were quantified with an assay from Biomedica.

Biomarkers of improved health after quitting smoking

Anti-Oxidized LDL Autoantibodies

Oxidized low-densitity lipoprotein (oxLDL) is believed to play a crucial role in in the development and progression of coronary artery disease (CAD) that involves atherosclerosis and inflammation. oxLDL in macrophages and smooth muscle cells accumulates and causes foam cell formation, which represents an early stage in the development of the disease. In fact, oxidized LDL appears to be around 24% higher in smokers compared to non-smokers (3). Autoantibodies targeting 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)

How can “Anti-Oxidized LDL Autoantibodies” be measured?

Autoantibodies targeting oxidatively modified LDL particles can easily be measured with a conventional ELISA assay.

Anti-Oxidized LDL Autoantibody ELISA (oLAB) assay (cat. no. BI-20032)

Highlights

• Widely cited
• Results in 2,5 hours
• 2 controls included
• Developed and manufactured by BIOMEDICA

 

Literature

1. Changes in biomarkers of endothelial function, oxidative stress, inflammation and lipids after smoking cessation: A cohort study. Colsoul ML et al., Eur J Clin Invest. 2023 Aug;53(8):e13996. doi: 10.1111/eci.13996. Epub 2023 Apr 8. PMID: 37000021.
2. Suitability of biomarkers of biological effects (BOBEs) for assessing the likelihood of reducing the tobacco related disease risk by new and innovative tobacco products: A literature review. Scherer G Regul Toxicol Pharmacol. 2018 Apr;94:203-233. doi: 10.1016/j.yrtph.2018.02.002. Epub 2018 Feb 9. PMID: 29428304.
3. Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou E et al., Free Radic Res. 2005 Apr;39(4):419-29. doi: 10.1080/10715760500072156. PMID: 16028367.
4. Biomarkers of improved health outcomes after smoking cessation. Neal L. Benowitz et al., Addiction Neuroscience,5: 2023, 100054,ISSN 2772-392. ISSN 2772-3925.

LRG – a reliable serum marker of disease activity in Inflammatory Bowel Disease: Inflammatory Bowel disease (IBD) like Crohn´s disease and ulcerative colitis affects millions of people worldwide. IBD is characterized by a chronic intestinal inflammation causing pain and swelling. The exact cause of IBD is not yet fully understood but it may involve the immune system, as well as genetic, and environmental factors. Research has shown that the immune system attacks harmless microorganisms or food in the gut that causes inflammation that leads to bowel injury.

Currently, the gold standard for assessing IBD disease activity are imaging tests, in particular endoscopy. The use of non-invasive biomarkers could be useful in clinical practice to monitor disease activity and to predict mucosal healing. CRP is often been used to evaluate IBD activity but research has shown that its specificity and low cut off value may not be ideal for use in clinical practice (1). Alternative biomarkers that more likely reflect intestinal inflammation are under investigation.

LRG – a reliable serum marker of disease activity in Inflammatory Bowel Disease

Leucine-rich α2 glycoprotein (LRG) is a protein that is produced by hepatocytes, neutrophils, and macrophages. It is a biomarker that is stimulated by inflammatory cytokines e.g. TNF-alpha, IL-1 beta, IL-11 and IL-6. However, studies have shown that release of LRG into the circulation does not depend on IL-6 and thus has a different mechanism from that of CRP. Therefore, LRG may better reflect intestinal inflammation better than LRG (3).

Recent studies have demonstrated that LRG is useful for predicting  mucosal healing and is a reliable serum biomarker for disease evaluation of clinical and endoscopic disease activity in individuals with inflammatory bowel disease (4-6).  

HOW CAN LRG BE MEASURED?

Leucine-rich α2 glycoprotein (LRG) can  easily and reliably be measured  in serum, plasma, urine and cell culture samples with a conventional ELISA (enzyme-linked immunosorbant assay) assay.

 

LRG ELISA – 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

 

Organizations for the study of IBD

• International Organization For the Study of Inflammatory Bowel Disease
• European Crohn’s and Colitis Organisation (ECCO)
• World Gastroenterology Organisation

 

Cited Literature

1. Leucine-rich alpha-2 glycoprotein as a marker of mucosal healing in inflammatory bowel disease. Yasutomi E, Inokuchi T, Hiraoka S, Takei K, Igawa S, Yamamoto S, Ohmori M, Oka S, Yamasaki Y, Kinugasa H, Takahara M, Harada K, Furukawa M, Itoshima K, Okada K, Otsuka F, Tanaka T, Mitsuhashi T, Kato J, Okada H. Sci Rep. 2021; 27;11(1):11086.
2.  Leucine-Rich Alpha-2 Glycoprotein Is a Reliable Serum Biomarker for Evaluating Clinical and Endoscopic Disease Activity in Inflammatory Bowel Disease. Shimoyama T, Yamamoto T, Yoshiyama S, Nishikawa R, Umegae S. Inflamm Bowel Dis. 2023; 29(9):1399-1408. 
3. Positioning and Usefulness of Biomarkers in Inflammatory Bowel Disease. Sakurai T, Saruta M. Digestion. 2023;104(1):30-41. 
4. Usefulness of Serum Leucine-rich Alpha 2 Glycoprotein in Crohn’s Disease: Is There Any Difference between Small Intestine and Colonic Lesions? Matsumoto S et al., Crohns Colitis 360. 2023; 23; 5(3):otad028.  Conclusions:  “LRG is more useful than CRP for predicting mucosal healing in patients with type L1. The superiority of LRG to CRP differs between small intestinal and colonic lesions.”
5. Leucine-Rich Alpha-2 Glycoprotein Is a Reliable Serum Biomarker for Evaluating Clinical and Endoscopic Disease Activity in Inflammatory Bowel Disease. Shimoyama T, Yamamoto T, Yoshiyama S, Nishikawa R, Umegae S.. Inflamm Bowel Dis. 2023; 29(9):1399-1408.  Conclusions: “Leucine-rich alpha-2 glycoprotein is a reliable serum biomarker for the assessment of clinical and endoscopic disease activity in patients with IBD. It can be an alternative to CRP for the assessment of UC.”
6. Leucine-rich alpha-2 glycoprotein is a potential biomarker to monitor disease activity in inflammatory bowel disease receiving adalimumab: PLANET study Shinzaki S et al., J Gastroenterol. 2021; 56(6):560-569. Conclusions: “Serum LRG is a useful biomarker of endoscopic activity both in CD and UC during the adalimumab treatment.”

 

Testing Cell Viability in Breast Cancer Cells with EZ4U – MTT Assay: In search for novel and effective drugs that don’t induce resistance or toxic effects the BIOMEDICA EZ4U – MTT Cell Viabilty Assay was used investigating various protein kinase C isoforms of Royleanones on the activity of breast cancer cell lines.

Royleanosones derive from plants from the genus Plectranthus with different varieties found throughout tropical Africa, Asia, and Australia. Various cytotoxic compounds with notable activities have been isolated from this plant. Learn more: Activity to Breast Cancer Cell Lines of Different Malignancy and Predicted Interaction with Protein Kinase C Isoforms of Royleanones.

Testing Cell Viability in Breast Cancer Cells with EZ4U – MTT Assay

In this study three natural Royleanones were tested for their inhibitory effects in three breast cancer cell lines namely: MCF-7 (high expression of estrogen receptors), SkBr3 (high expression of Her2^NEU ), and SUM159 (negative for both hormone receptors and for Her2).

In brief: the cell lines were grown at 37°C with 5% CO2. To test cell viability, cells were seeded at 10.000 cells per well and left for 24h. After cell attachment they were treated with various concentrations of the three natural Royleanones compounds and incubated for 24h. Cell viability was assayed with the EZ4U – MTT assay (Biomedica, Austria).

Short description: 20µl of the EZ4U substance (colorless non-toxic tetrazolium salts) was 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)

 

• Non-radioactive & non-toxic assay
• Reliable & Sensitive
• Convenient single-step incubation  – for use on living cells
• Widely cited in more than 250 publications

 

Find out more: 

• BROCHURE – EZ4U cell proliferation and cytotoxicity assay.
• Protocol Booklet

 

Citations using the BIOMEDICA EZ4U Assay (cat. no. BI-5000) in various cell-lines

Study on Anticancer Activity of 4, 4′-[1,4-phenylenebis(1,3,4-thiadiazole-5,2-diyl)] bis (azaneylylidene) bis (methaneylylidene) diphenolon Breast Cancer Cells. Saeed BMS et al., Arch Razi Inst. 2021; 76(4):821-827. 

Aberrant DNA Methylation, Expression, and Occurrence of Transcript Variants of the ABC Transporter ABCA7 in Breast Cancer. Zappe K et al., Cells. 2023;  24;12(11):1462. 

AQP3-Dependent PI3K/Akt Modulation in Breast Cancer Cells. Mlinarić M et al., Int J Mol Sci. 2023; 1;24(9):8133. 

Identification of F-Box/SPRY Domain-Containing Protein 1 (FBXO45) as a Prognostic Biomarker for TMPRSS2-ERG-Positive Primary Prostate Cancers. von Danwitz M et al.,  Cancers (Basel). 2023; 21;15(6):1890. 

Dual targeting of BCL-2 and MCL-1 in the presence of BAX breaks venetoclax resistance in human small cell lung cancer. Valko Z et al., Br J Cancer. 2023; 128(10):1850-1861. 

for more citations click here 

 

 

Periostin is a soluble extracellular matrix protein that plays a role in bone formation and has been linked to the development of various cancers, including breast cancer. It is secreted in the tumor environment mostly by cancer associated fibroblasts promoting the progression of cancer which includes cell survival, metastasis and also chemoresistance.

Periostin – a prognostic biomarker in breast cancer

The prognostic value of serum Periostin was investigated in a cohort of 509 non metastatic breast cancer patients. The results demonstrated the importance of Periostin in breast cancer and showed that serum Periostin is a prospective indicator for disease prognosis, regardless of the existence of micrometastases. Read more: High serum levels of periostin are associated with a poor survival in breast cancer.

Human serum Periostin levels were measured with the BIOMEDICA PERIOSTIN  ELISA.

Periostin ELISA (cat. no. BI-20433)

• • Low sample volume – 10 µl sample / well
  • For human serum/plasma/cell culture supernatants
  • High quality assay – validation following international guidelines
  • All reagents included – 7 ready to use calibrators and 2 controls
  • Numerous citations using the Biomedica Periostin ELISA

 

example of a BIOMEDICA ELISA assay kit with color-coded reagents, ready to use pre-diluted calibrators/standards and 2 (high/low) kit controls

 

The full characterization of the Periostin ELISA assay has been published in:

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. doi: 10.1002/jcla.22252. 

Also available: Mouse Periostin ELISA (cat. no. BI-20433MS)

• Low sample volume – ≤ 5 µl / well
• For serum/plasma/cell culture supernatants

 

All ELISA kits are developed and manufactured by BIOMEDICA-Vienna/Austria

Related publications

High serum levels of periostin are associated with a poor survival in breast cancer. Rachner TD, Göbel A, Hoffmann O, Erdmann K, Kasimir-Bauer S, Breining D, Kimmig R, Hofbauer LC, Bittner AK. Breast Cancer Res Treat. 2020 Apr;180(2):515-524. doi: 10.1007/s10549-020-05570-0. Epub 2020 Feb 10. PMID: 32040688.

Periostin gene expression in neu-positive breast cancer cells is regulated by a FGFR signaling cross talk with TGFβ/PI3K/AKT pathways. Labrèche C, Cook DP, Abou-Hamad J, Pascoal J, Pryce BR, Al-Zahrani KN, Sabourin LA. Breast Cancer Res. 2021 Nov 22;23(1):107. doi: 10.1186/s13058-021-01487-8. PMID: 34809697; PMCID: PMC8607680.

Periostin Exon-21 Antibody Neutralization of Triple-Negative Breast Cancer Cell-Derived Periostin Regulates Tumor-Associated Macrophage Polarization and Angiogenesis. Fujikawa T, Sanada F, Taniyama Y, Shibata K, Katsuragi N, Koibuchi N, Akazawa K, Kanemoto Y, Kuroyanagi H, Shimazu K, Rakugi H, Morishita R.Cancers (Basel). 2021 Oct 11;13(20):5072. doi: 10.3390/cancers13205072. PMID: 34680221; PMCID: PMC8533925.

The Role of Extracellular Matrix Proteins in Breast Cancer. Lepucki A, Orlińska K, Mielczarek-Palacz A, Kabut J, Olczyk P, Komosińska-Vassev K. J Clin Med. 2022 Feb 25;11(5):1250. doi: 10.3390/jcm11051250. PMID: 35268340; PMCID: PMC8911242.

Fibroblast growth factor 23 (FGF23) is a protein that is produced by osteocytes, which are specialized cells found in bone tissue (1). FGF23 plays a key role in maintaining the balance of phosphate and vitamin D in the body (2). FGF23 primary function is to regulate blood phosphate levels by stimulating the removal of excess phosphate by the kidneys.

Elevated FGF23 levels in the bloodstream lead to a reduction in blood phosphate levels, a decrease in 1,25(OH)2 vitamin D, and the development of osteomalacia (3).

In addition to its known effects on bone and mineral regulation, it has been suggested that FGF23 may play a significant role in the development of type 2 diabetes mellitus (4). Findings showed that FGF23 is linked to indicators of obesity, metabolic syndrome and insulin levels.

FGF23 associated with the development of gestational diabetes

Gestational diabetes mellitus (GDM) is a type of diabetes that can develop during pregnancy in women who do not have existing diabetes. It is defined as glucose intolerance that is recognized first during pregnancy. GDM and impaired glucose tolerance during pregnancy are associated with persistent metabolic dysfunction three years after delivery, independent of other clinical risk factors (5).

FGF23 associated with the development of gestational diabetes

In the current study by Hocher CF and colleagues (6), the researchers investigated the association of maternal and foetal FGF23 with gestational diabetes mellitus (GDM) in a German birth cohort. Both FGF23 (c-terminal) and FGF23 intact levels were analysed in 826 random mother/child pairs with ELISA assays from BIOMEDICA. Of note: c-terminal FGF23 ELISA kits recognize both full-length intact FGF23 and c-terminal fragments of FGF23, and intact FGF23 assays solely detect full-length intact FGF23. The use of both assays in the current study is based on evidence from head to head comparison studies where clinical associations were reported based on the FGF23 assays used ( 7, 8). In summary, the study of a representative German birth cohort showed that maternal but not foetal iFGF23 is independently associated with GDM (6).

FGF23 can reliably be measured by ELISA assay

FGF23 intact ELISA, cat. no. BI-20700

FGF23 (c-terminal) ELISA,  cat. no.  BI-20702

BIOMEDICA´s FGF23 ELISA Kits:

• Low sample volume: 50µl / well
• For plasma (EDTA, citrate, heparın), serum, urine, cell culture supernatant
• Kit controls (high and low) included
• Full validation following international quality guidelines
• Numerous top journal citations

 

 

Literature:

1. The osteocyte: A multifunctional cell within the bone. Tresguerres FGF, Torres J, López-Quiles J, Hernández G, Vega JA, Tresguerres IF. Ann Anat. 2020 Jan;227:151422. doi: 10.1016/j.aanat.2019.151422. Epub 2019 Sep 26. Erratum in: Ann Anat. 2020 Jul;230:151510. PMID: 31563568.
2. Role of phosphate sensing in bone and mineral metabolism. Chande S, Bergwitz C. Nat Rev Endocrinol. 2018 Nov;14(11):637-655. doi: 10.1038/s41574-018-0076-3. PMID: 30218014; PMCID: PMC8607960.
3. FGF23 and Hypophosphatemic Rickets/Osteomalacia. Takashi Y, Kawanami D, Fukumoto S. Curr Osteoporos Rep. 2021 Dec;19(6):669-675. doi: 10.1007/s11914-021-00709-4. Epub 2021 Nov 10. PMID: 34755323.
4. The Emerging Role of Bone-Derived Hormones in Diabetes Mellitus and Diabetic Kidney Disease. Li Y, Gu Z, Wang J, Wang Y, Chen X, Dong B. Front Endocrinol (Lausanne). 2022 Jul 11;13:938830. doi: 10.3389/fendo.2022.938830. PMID: 35966090; PMCID: PMC9367194.
5. Gestational glucose tolerance and maternal metabolic profile at 3 years postpartum. Stuebe AM, Mantzoros C, Kleinman K, Gillman MW, Rifas-Shiman S, Seely EW, Rich-Edwards J. Obstet Gynecol. 2011 Nov;118(5):1065-1073. doi: 10.1097/AOG.0b013e3182325f5a. PMID: 22015874; PMCID: PMC3268071.
6. Fibroblast growth factor 23 is associated with the development of gestational diabetes mellitus. Hocher CF, Chen X, Zuo J, Horvathova K, Hocher B, Krämer BK, Chu C. Diabetes Metab Res Rev. 2023 Aug 8:e3704. doi: 10.1002/dmrr.3704. Epub ahead of print. PMID: 37553983.
7. C-terminal and intact FGF23 in kidney transplant recipients and their associations with overall graft survival. Chu C, Elitok S, Zeng S, Xiong Y, Hocher CF, Hasan AA, Krämer BK, Hocher B. BMC Nephrol. 2021 Apr 8;22(1):125. doi: 10.1186/s12882-021-02329-7. PMID: 33832449; PMCID: PMC8033679.
8. Fibroblast growth factor 23 concentrations and modifying factors in children from age 12 to 24 months. Enlund-Cerullo M, Hauta-Alus H, Valkama S, Rosendahl J, Andersson S, Mäkitie O, Holmlund-Suila E. Bone. 2020 Dec;141:115629. doi: 10.1016/j.bone.2020.115629. Epub 2020 Sep 10. PMID: 32919110.

 

 

Breast cancer is the most common detected cancer worldwide with 2.3 million new cases and 685, 000 deaths in 2020 (1).

1 in 8 women are predicted to be diagnosed with breast cancer in their lifetime. 

Early detection of breast cancer is the key to successful outcomes.

Breast Cancer Awareness Month – October 2023

Find out more about some of the different breast cancer associations worldwide and learn about what you should be aware of:

• National Breast Cancer Foundation Inc
• WHO Europe
• Breast Cancer UK
• Breast Cancer Foundation New Zealand
• Canadian Cancer Society
• Africa Health Organisation
• American Association for Cancer Research
• Japanese Breast Cancer Society

 

BIOMEDICA – Biomarker ELISA Kits for Breast Cancer Research

PERIOSTIN ▪ NEUROPILIN-1 ▪ OSTEPROTEGERIN ▪ soluble RANKL ▪ SEMAPHORIN 4D

check out our Brochure Biomarker ELISAs in Oncology

Assay Kit Highlights

√  EASY –ready to use calibrators & controls included (color-coded reagents)
√  FULL VALIDATION PACKAGE – assays are optimized for clinical samples
√   HIGH QUALITY GUARANTEED – results you can rely on
√  WIDELY CITED in 1500 + publications
√  COMPETENT CUSTOMER SERVICE

 

BIOMEDICA – Complete ready-to-use ELISA kits for superior performance and reproducibility

Literature

1. Current and future burden of breast cancer: Global statistics for 2020 and 2040. Arnold M, Morgan E, Rumgay H, Mafra A, Singh D, Laversanne M, Vignat J, Gralow JR, Cardoso F, Siesling S, Soerjomataram I Breast. 2022 Dec;66:15-23. doi: 10.1016/j.breast.2022.08.010. Epub 2022 Sep 2. PMID: 36084384; PMCID: PMC9465273.
2. The prevention and treatment of breast cancer- related lymphedema: A review. McEvoy MP, Gomberawalla A, Smith M, Boccardo FM, Holmes D, Djohan R, Thiruchelvam P, Klimberg S, Dietz J, Feldman S. Front Oncol. 2022 Dec 6;12:1062472. doi: 10.3389/fonc.2022.1062472. PMID: 36561522; PMCID: PMC9763870.
3. Breast Cancer in India: Screening, Detection, and Management. Bhardwaj PV, Dulala R, Rajappa S, Loke C. Hematol Oncol Clin North Am. 2023 Jun 15:S0889-8588(23)00069-2. doi: 10.1016/j.hoc.2023.05.014. Epub ahead of print. PMID: 37330342.
4. Outcomes in breast cancer-does ethnicity matter? Yap YS ESMO Open. 2023 Jun;8(3):101564. doi: 10.1016/j.esmoop.2023.101564. Epub 2023 Jun 7. PMID: 37290358; PMCID: PMC10265619.

 

 

Ovarian cancer ranks as the primary contributor of death in patients with gynecological malignancies and over 70% of patients receive their diagnosis at an advanced stage.  Despite advances in treatment, a large number of ovarian cancer patients still are confronted with a poor overall prognosis. The discovery of blood-based biomarkers that can predict outcomes or provide prognostic insights is promising as findings could lead to the identification of novel drug targets and tailored treatments for ovarian cancer patients.

Neuropilin-1 is a blood-based prognostic biomarker in patients with ovarian cancer

Neuropilin-1 is a transmembrane cell-surface receptor that has been implicated in various aspects of cancer biology. It is found on the surface of cancer cells and is shed into the circulation where it can successfully be measured in blood samples with a conventional ELISA assay. Neuropilin-1 is associated with tumor progression and the formation of new blood vessels to support tumor growth is a process called angiogenesis. Neuropilin-1 interacts with molecules like vascular endothelial growth factor (VEGF) and Semaphorins promoting the survival and migration of cancer cells.

Neuropilin-1 is a blood-based prognostic biomarker in patients with ovarian cancer

The BIOMEDICA Neuropilin-1 (NRP-1) ELISA assay was highlighted in a recent study (1) showing for the first time that NRP-1 is a blood based prognostic biomarker in patients with ovarian cancer. The results encourage further research to analyse whether soluble NRP-1 could be considered as a predictive and prognostic routine tool in patients with ovarian cancer.

EASY measurement of soluble Neuropilin-1 in blood samples:

Total Soluble Neuropilin-1 ELISA | BI-20409

• Method: conventional 96-well ELISA format
• Sample volume: 10µl / well
• Sample matrix: human serum, plasma, urine, cell culture supernatant
• High homology between human NRP-1 and non-human samples –  protocol available
• Full validation package – click here 

 

Only assay that measures free and ligand bound soluble NRP1

Reference

1. Clinical impact of soluble Neuropilin-1 in ovarian cancer patients and its association with its circulating ligands of the HGF/c-MET axis.   Klotz DM, Kuhlmann JD, Link T, Goeckenjan M, Hofbauer LC, Göbel A, Rachner TD, Wimberger P. Front Oncol. 2022 Oct 21;12:974885. doi: 10.3389/fonc.2022.974885. PMID: 36338759; PMCID: PMC9635484.

 

Abstract

Background: Neuropilin (NRP) is a transmembrane protein, which has been shown to be a pro-angiogenic mediator and implicated as a potential driver of cancer progression. NRP-1 up-regulation in ovarian cancer tissue predicts poor prognosis. However, the clinical relevance of the soluble form of NRP-1 (sNRP-1) as a circulating biomarker in ovarian cancer patients is unknown.

Methods/patients cohort: sNRP-1 levels were quantified in a cohort of 88 clinically documented ovarian cancer patients by a commercially available sNRP-1 enzyme-linked immunosorbent assay (ELISA) kit (Biomedica, Vienna, Austria). Patients (81.8% with FIGOIII/IV) received primary cytoreductive surgery with the aim of macroscopic complete resection (achieved in 55.7% of patients) and the recommendation of adjuvant chemotherapy in line with national guidelines.

Results: Higher levels of sNRP-1 reflected more advanced disease (FIGO III/IV) and indicated a trend towards suboptimal surgical outcome, i.e. any residual tumor. sNRP-1 was neither related to the patients’ age nor the BRCA1/2 mutational status. Patients with higher sNRP-1 levels at primary diagnosis had a significantly reduced progression-free survival (PFS) (HR = 0.541, 95%CI: 0.304 – 0.963; p = 0.037) and overall survival (OS) (HR = 0.459, 95%CI: 0.225 – 0.936; p = 0.032). Principal component analysis showed that sNRP-1 levels were unrelated to the circulating hepatocyte growth factor (HGF) and the soluble ectodomain of its receptor the tyrosine kinase mesenchymal-epithelial transition (c-MET), suggesting that there is no proportional serological concentration gradient of soluble components of the NRP-1/HGF/c-MET signaling axis.

Conclusions: In line with the previously shown tissue-based prognostic role, we demonstrated for the first time that sNRP-1 can also act as a readily accessible, prognostic biomarker in the circulation of patients with ovarian cancer at primary diagnosis. Given its known role in angiogenesis and conferring resistance to the poly ADP-ribose polymerase (PARP) inhibitor olaparib in vitro, our results encourage more detailed investigation into sNRP-1 as a potential predictive biomarker for bevacizumab and/or PARP-inhibitor treatment.

 

 

 

 

We at BIOMEDICA develop and manufacture ELISA assays to reliably quantify cytokines in biological fluids.

Our ELISA kits are rigorously validated according to international quality standards to ensure their consistency, specificity, precision, and robustness.

All of the validation data can be found on the respective product pages on our website www.bmgrp.com . 

Quantify Cytokines with an ELISA Assay from BIOMEDICA

We designed our assays to quantify the cytokine of interest in serum, plasma or cell culture supernatants.

Standardized harmonization of results with WHO reference reagents

To ensure a harmonized standardization, WHO reference reagents / standards are employed for cross-comparison of results (1)

Quantify Cytokines with an ELISA Assay from BIOMEDICA

IL-6 ELISA (cat. no. BI-IL6)

The IL-6 (Interleukin-6) ELISA kit from BIOMEDICA is a conventional  sandwich ELISA kit. The assay incorporates two epitope-mapped antibodies that specifically bind to human IL-6 in the respective samples.

Capture antibody : is a  recombinant IL-6 antibody that is specific for human IL-6, which is pre-coated on a 96-well microtiter plate. 

Detection antibody: is a polyclonal IL-6 antibody (labeled with streptavidin-HRPO), which is specific for human IL-6.

• • Assay method: sandwich ELISA
  • Sample types: serum, plasma, cell-culture, urine
  • Sample volume: 100µl / well
  • Assay time: 4.5 hrs
  • Standard range: 0; 3.1 – 200 pg/ml
  • Sensitivity: LLOQ: 0.78 pg/ml (measurable concentrations in both serum AND plasma samples!),  LOD: 0.28 pg/ml
  • Precision: in-between-run (n=9 ): ≤ 6 % CV, within-run (n=3): ≤ 7 % CV
  • Specificity:  the ELISA recognizes endogenous (natural) and recombinant and human IL-6
  • Standard matrix: 8 x pre-diluted standards, lyophilized,  in a serum based matrix containing recombinant human Il-6
  • Controls: 1x high and 1x low control included in the kit

 

VEGF ELISA (cat. no. BI-VEGF)

The VEGF (Vascular Endothelial Growth Factor) ELISA kit from BIOMEDICA is a conventional sandwich ELISA kit. The assay incorporates two epitope-mapped antibodies that specifically bind to human VEGF in the respective samples.

Capture antibody is a recombinant VEGF antibody that is specific for human VEGF, which is pre-coated on a 96-well microtiter plate,  recognizing a structural epitope in the conserved receptor binding- site of the VEGF peptide. All VEGF isoforms are detected.

Detection antibody: is a polyclonal VEGF antibody (labeled with streptavidin-HRPO), which is specific for human VEGF. The antibody recognizes multiple linear epitopes that are concentrated in the first 120 amino acids in the N-terminal region of the VEGF molecule.   

• Assay method: sandwich ELISA
• Sample types: serum, plasma, cell-culture, urine
• Sample volume: 10µl / well
• Assay time: 4.5 hrs
• Standard range: 0; 31.2 – 2000 pg/ml
• Sensitivity: LLOQ: 15.6 pg/ml); LOD: 2.5 pg/ml
• Precision: in-between-run: (n=3): ≤ 6 % CV, Within-run (n=3): ≤ 3 % CV
• Specificity: the ELISA recognizes endogenous (natural) human VEGF including all circulating VEGF isoforms including VEGF165b and recombinant VEGF
• Standard matrix: 8 x vials of pre-diluted standards, lyophilized, in a serum based matrix containing recombinant human VEGF
• Controls: 2 controls (high and low) included in the kit

 

ANGIOPOIETIN-2 ELISA (cat. no. BI-ANG2)

The ANG2 (Angiopoietin-2) ELISA kit from BIOMEDICA  is a conventional sandwich ELISA kit. The assay incorporates two epitope-mapped antibodies that specifically bind to human Angiopoietin-2 in the respective samples.

Capture antibody is a recombinant ANG2 antibody that is specific for human VEGF, which is pre-coated on a 96-well microtiter plate. The capture antibody has a structural epitope that binds to the receptor binding site of human Angiopoietin-2. The receptor binds to TEL / TIE2 protein.

Detection antibody: is a polyclonal ANG2 antibody (labeled with streptavidin-HRPO), which is specific for human ANG2. The antibody recognizes several linear epitopes that are distributed over the entire Angiopoietin-2 molecule.  

• Assay method: sandwich ELISA
• Sample types: serum, plasma, cell-culture, urine
• Sample volume: 20µl / well
• Assay time: 3.5 hrs
• Standard range: 0; 0.68 – 21.96 ng/ml (= 0 ; 12.5 – 400 pmol /l  (conversion 1 pmol/l = 54.9 pg/ml (MW: 54.9 kDa)
• Sensitivity: LLOQ : 0.34 ng/ml (=6.3 pmol/l); LOD: 0.20 pg/ml (= 3.7 pmol/l)
• Precision: In-between-run (n=9): ≤ 6 % CV, Within-run (n=3): ≤ 8 % CV
• Specificity: the ELISA recognizes endogenous (natural) human ANG2 and recombinant ANG2.  Most probably all three ANG2 isoforms are detected by the assay, as determined by epitope mapping and analysis of the ANG2 sequence. This ELISA assay does not detect ANG1.
• Standard matrix: 8 x vials of pre-diluted standards, lyophilized, in a serum based matrix containing recombinant human ANG2.
• Controls: 2 controls (high and low) included in the kit

 

Mouse /rat Angiopoietin-2 ELISA (cat. no. BI-ANG2MR) also available

Related publications

1. International reference reagents
2. Angiopoietin-2 Promotes Pathological Angiogenesis and Is a Therapeutic Target in Murine Nonalcoholic Fatty Liver Disease. Lefere S, Van de Velde F, Hoorens A, Raevens S, Van Campenhout S, Vandierendonck A, Neyt S, Vandeghinste B, Vanhove C, Debbaut C, Verhelst X, Van Dorpe J, Van Steenkiste C, Casteleyn C, Lapauw B, Van Vlierberghe H, Geerts A, Devisscher L. Hepatology. 2019 Mar;69(3):1087-1104. doi: 10.1002/hep.30294. Epub 2019 Feb 12. PMID: 30259536.

The development of heart failure is a concerning complication for patients who undergo heart valve and/or coronary bypass grafting surgery. In this context, risk prediction tools for patients undergoing cardiac surgery are of great interest. This article highlights a recent study investigating the predictive power of preoperative FGF23 levels in patients undergoing cardiac surgery.

Fibroblast growth factor 23 (FGF23)

Fibroblast growth factor 23 (FGF23) is a protein that plays a vital role in regulating phosphate and vitamin D levels and is associated with an elevated cardiovascular risk. A recent study by Hofer F et al.,  aimed to explore how FGF23 affects cardiovascular outcomes in a group of patients who underwent cardiac surgery. The analysis included a cohort of 451 patients who were observed for a median duration of 3.9 years. The results indicated that individuals with higher FGF23 levels showed elevated incidence of cardiovascular death. Therefore, the assessment of FGF23 may be a tool for risk stratification and could allow early identification of patients who have a high risk for adverse cardiovascular outcomes.

“Plasma FGF‐23 concentrations were assessed with a sandwich enzyme immunoassay (Biomedica Medizinprodukte GmbH, Vienna, Austria). The intra‐assay and interassay precision values were ≤12% and ≤10%, respectively. The assay limit of detection (0 pmol/L+3 SDs) was 0.08 pmol/L. For better clinical evaluation, FGF‐23 was converted to units of pg/mL, with a conversion factor of 1 pg/mL=0.133 pmol/L.”

Predictive power of preoperative FGF23 levels in patients undergoing cardiac surgery

Routine measurement of preoperative FGF23 levels may improve the detection of high-risk patients undergoing cardiac surgery.

 BIOMEDICA  FGF23 ELISA assays for  serum & plasma samples

FGF23 (c-terminal) ELISA (cat. no BI-20702) and FGF23 intact ELISA (cat. no.  BI-20700)

• developed & manufactured by Biomedica , Vienna, Austria
• fully validated according to international quality guidelines
• numerous citations in top journals

 

Related Publications

Relationship of Fibroblast Growth Factor 23 With Hospitalization for Heart Failure and Cardiovascular Outcomes in Patients Undergoing Cardiac Surgery. Hofer F, Hammer A, Pailer U, Koller L, Kazem N, Steinacher E, Steinlechner B, Andreas M, Laufer G, Wojta J, Zelniker TA, Hengstenberg C, Niessner A, Sulzgruber P. J Am Heart Assoc. 2023 Mar 7;12(5):e027875. doi: 10.1161/JAHA.122.027875. Epub 2023 Feb 21. PMID: 36802737; PMCID: PMC10111457.

Abstract

Background Fibroblast growth factor 23 (FGF-23) is crucial in regulating phosphate and vitamin D metabolism and is moreover associated with an increased cardiovascular risk. The specific objective of this study was to investigate the influence of FGF-23 on cardiovascular outcomes, including hospitalization for heart failure (HHF), postoperative atrial fibrillation, and cardiovascular death, in an unselected patient population after cardiac surgery. Methods and Results Patients undergoing elective coronary artery bypass graft and/or cardiac valve surgery were prospectively enrolled. FGF-23 blood plasma concentrations were assessed before surgery. A composite of cardiovascular death/HHF was chosen as primary end point. A total of 451 patients (median age 70 years; 28.8% female) were included in the present analysis and followed over a median of 3.9 years. Individuals with higher FGF-23 quartiles showed elevated incidence rates of the composite of cardiovascular death/HHF (quartile 1, 7.1%; quartile 2, 8.6%; quartile 3, 15.1%; and quartile 4, 34.3%). After multivariable adjustment, FGF-23 modeled as a continuous variable (adjusted hazard ratio for a 1-unit increase in standardized log-transformed biomarker, 1.82 [95% CI, 1.34-2.46]) as well as using predefined risk groups and quartiles remained independently associated with the risk of cardiovascular death/HHF and the secondary outcomes, including postoperative atrial fibrillation. Reclassification analysis indicated that the addition of FGF-23 to N-terminal pro-B-type natriuretic peptide provides a significant improvement in risk discrimination (net reclassification improvement at the event rate, 0.58 [95% CI, 0.34-0.81]; P<0.001; integrated discrimination increment, 0.03 [95% CI, 0.01-0.05]; P<0.001). Conclusions FGF-23 is an independent predictor of cardiovascular death/HHF and postoperative atrial fibrillation in individuals undergoing cardiac surgery. Considering an individualized risk assessment, routine preoperative FGF-23 evaluation may improve detection of high-risk patients.

A single preoperative FGF23 measurement is a strong predictor of outcome in patients undergoing elective cardiac surgery: a prospective observational study. Speer T, Groesdonk HV, Zapf B, Buescher V, Beyse M, Duerr L, Gewert S, Krauss P, Poppleton A, Wagenpfeil S, Fliser D, Schaefers HJ, Klingele M.Crit Care. 2015 Apr 23;19(1):190. doi: 10.1186/s13054-015-0925-6. PMID: 25902817; PMCID: PMC4424828.

Medication-related osteonecrosis of the jaw (MRONJ) is a severe side effect that can occur in patients taking medications often used for treating cancer and osteoporosis. MRONJ occurs when blood supply to a part of the bone is disrupted. This leads to a gradual destruction of bone in the jaw.

Semaphorin 4D (SEMA4D) is a protein that has various important functions in both the immune- and the nervous system. SEMA4D also plays a role in bone biology as it influences bone remodeling by interacting between immune and bone cells, specifically osteoclasts, the cells responsible for breaking down bone.

SEMAPHORIN 4D in medication-related osteonecrosis of the jaw

In a recent study researchers investigated the clinical significance of circulating SEMA4D in the early diagnosis of MRONJ:

Clinical values of serum Semaphorin 4D (Sema4D) in medication‑related osteonecrosis of the jaw. 

Semaphorin 4D can easily be measured in blood samples with an ELISA assay

Biomedica offers a high-quality test kit to measure soluble SEMA4D in human plasma samples

Semaphorin 4D (SEMA4D) ELISA Assay Highlights:

• • Only Sema4D ELISA assay that is fully validated
  • 10 µl / well sample volume – sample predilution is not required
  • Available reference values for healthy individuals

To see the protocol booklet for SEMA4D ELISA just click here

Related literature

 Clinical values of serum Semaphorin 4D (Sema4D) in medication‑related osteonecrosis of the jaw. Mu H, Pang Y, Liu L, Liu J, Liu C. Eur J Med Res. 2023 Mar 30;28(1):140. doi: 10.1186/s40001-023-01095-6. PMID: 36998031; PMCID: PMC10061851.

Abstract

Background: Bisphosphonates (BPs) are widely used in clinical practice to prevent and treat bone metabolism-related diseases. Medication-related osteonecrosis of the jaw (MRONJ) is one of the major sequelae of BPs use. Early prediction and intervention of MRONJ are of great significance.

Methods: Ninety-seven patients currently on treatment with BPs or with a history of BPs usage and 45 healthy volunteers undergoing dentoalveolar surgery were included in this study. Participants’ serum Semaphorin 4D (Sema4D) levels were measured and analyzed before participants underwent surgery (T0) and after a 12-month follow-up (T1). Kruskal-Wallis test and ROC analysis were used to examine the predictive effect of Sema4D on MRONJ.

Results: Sema4D levels in serum of patients corresponding to confirmed MRONJ were significantly lower at both T0 and T1 time points compared to non-MRONJ and healthy controls. Sema4D has a statistically predictive effect on the occurrence and diagnosis of MRONJ. Serum Sema4D levels were significantly reduced in MRONJ class 3 patients. MRONJ patients who received intravenous BPs had significantly lower Sema4D levels than those who received oral BPs.

Conclusion: Serum Sema4D level has predictive value for the onset of MRONJ in BPs users within 12 weeks after dentoalveolar surgery

Interventions for managing medication-related osteonecrosis of the jaw. Beth-Tasdogan NH, Mayer B, Hussein H, Zolk O. Cochrane Database Syst Rev. 2017 Oct 6;10(10):CD012432. doi: 10.1002/14651858.CD012432.pub2. Update in: Cochrane Database Syst Rev. 2022 Jul 12;7:CD012432. PMID: 28983908; PMCID: PMC6485859.

Possible pathogenic engagement of soluble Semaphorin 4D produced by γδT cells in medication-related osteonecrosis of the jaw (MRONJ). Movila A, Mawardi H, Nishimura K, Kiyama T, Egashira K, Kim JY, Villa A, Sasaki H, Woo SB, Kawai T. Biochem Biophys Res Commun. 2016 Nov 4;480(1):42-47. doi: 10.1016/j.bbrc.2016.10.012. Epub 2016 Oct 5. PMID: 27720716.

 

.

With advancing age, the human body´s natural capacity to consistently renew bones and maintain their resilience diminishes. This decline is enhanced by conditions like osteoporosis. This poses a significant health concern for the elderly and is becoming a growing economic challenge on society. In an effort to address this problem, scientists are actively seeking novel therapeutic strategies to enhance bone regeneration.

 

Building bone with bioinspired molecules

By employing computer models and simulations, a Dresden-based team created innovative bio-inspired compounds that enhance bone regeneration in mice. These compounds can be incorporated into biomaterials, allowing for their localized introduction into bone defects. These newly developed molecules derive from glycosaminoglycans, which are extended sugar chains like hyaluronic acid or heparin. These molecules could be used to turn-off proteins like DKK-1 and Sclerostin that block bone regeneration that could lead to new and more effective therapies for bone diseases.

DKK-1 and SCLEROSTIN are two important proteins that play significant roles in regulating bone health and development.

DKK-1 (Dickkopf-1) acts as an inhibitor of the Wnt signaling pathway. This pathway is crucial for bone formation. Overexpression of DKK-1 leads to a decrease in bone formation characterized by conditions with impaired bone density and increased fragility, such as osteoporosis.

SCLEROSTIN (SOST) is a Wnt signaling inhibitor and a negative regulator of bone formation. Sclerostin is primarily produced by osteocytes, which are bone cells embedded within the bone matrix.

Sclerostin and DKK-1 can be measured in human serum samples with an ELISA assay.

 

Biomedica´s DKK-1 ELISA assay (cat. no. BI-20413)

• 20µl sample volume
• Widely cited
• No sample dilution
• Fully validated

 

Biomedica´s Sclerostin ELISA assay (cat. no. 20492)

• The internationally most referenced Sclerostin ELISA!
• Rigorously validated according to FDA/ICH/EMEA guidelines
• 20µl sample volume

 

also available: Bioactive Sclerostin ELISA (cat. no. BI-20472)

Rational engineering of glycosaminoglycan-based Dickkopf-1 scavengers to improve bone regeneration. Ruiz-Gómez G, Salbach-Hirsch J, Dürig JN, Köhler L, Balamurugan K, Rother S, Heidig SL, Moeller S, Schnabelrauch M, Furesi G, Pählig S, Guillem-Gloria PM, Hofbauer C, Hintze V, Pisabarro MT, Rademann J, Hofbauer LC. Biomaterials. 2023 Jun;297:122105. doi: 10.1016/j.biomaterials.2023.122105. Epub 2023 Mar 31. PMID: 37031548.

Abstract

The WNT signaling pathway is a central regulator of bone development and regeneration. Functional alterations of WNT ligands and inhibitors are associated with a variety of bone diseases that affect bone fragility and result in a high medical and socioeconomic burden. Hence, this cellular pathway has emerged as a novel target for bone-protective therapies, e.g. in osteoporosis. Here, we investigated glycosaminoglycan (GAG) recognition by Dickkopf-1 (DKK1), a potent endogenous WNT inhibitor, and the underlying functional implications in order to develop WNT signaling regulators. In a multidisciplinary approach we applied in silico structure-based de novo design strategies and molecular dynamics simulations combined with synthetic chemistry and surface plasmon resonance spectroscopy to Rationally Engineer oligomeric Glycosaminoglycan derivatives (_REGAG) with improved neutralizing properties for DKK1. In vitro and in vivo assays show that the GAG modification to obtain _REGAG translated into increased WNT pathway activity and improved bone regeneration in a mouse calvaria defect model with critical size bone lesions. Importantly, the developed _REGAG outperformed polymeric high-sulfated hyaluronan (sHA3) in enhancing bone healing up to 50% due to their improved DKK1 binding properties. Thus, rationally engineered GAG variants may represent an innovative strategy to develop novel therapeutic approaches for regenerative medicine

ELISA – Luminex – NGS – microRNA

Analytical Service Measurements – we offer tailored analytical testing services for

ELISA  & Luminex Assays, NGS & microRNA  and can support your research with:

 

• Expertise: experienced laboratory staff
• Quality: highest quality equipment
• Flexibility: tailored solutions according to your project needs and budget
• Speed: rapid turn-around time to meet your deadlines
• Results: verified and comprehensive results presented in an analytical report

Learn more about our ELISA and Luminex Workflow chart and our microRNA service details click here

Analytical Service Measurements – Biomedica

ELISA

We will be happy to measure any kind of biomarker of your choice with an ELISA assay (enzyme linked immunosorbent assay) including our propriety Biomedica biomarker ELISA kits (www.bmgrp.com).

Our Service Measurement Applications include analysis of samples for:

• • Clinical studies (serum and plasma samples)
  • Pre-clinical studies (any species)

 

Of note: Biomedica offers several biomarker ELISA kits for the application in various animal models: Rat-NT-proBNP ELISA and  NT-proANP ELISA (widely used in rat and mice,  works also in also in rabbit or pig models).

• • Custom applications – Learn about our workflow

RNA Services

Together with TamiRNA we offer a range of quality RNA services:

• RNA extraction: Biofluids including serum, plasma, and extracellular vesicles. Cells and tissues – the quality control of total RNA is carried out utilizing bioanalyser chips. 
• Next-generation sequencing- NGS.
• RT-qPCR
• We also analyse cell-type specific microRNA/mRNA  in complex tissues and offer custom analysis of microRNA signatures.
• Finally, we carry out the analysis of established microRNA signatures to predict fracture risk, platelet function, and toxicity.

 

Further reading

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.

RNA Extraction from Cartilage: Issues, Methods, Tips. Pagani S, Maglio M, Sicuro L, Fini M, Giavaresi G, Brogini S. Int J Mol Sci. 2023 Jan 20;24(3):2120. doi: 10.3390/ijms24032120. PMID: 36768444; PMCID: PMC9917073.

A Comprehensive Review of Performance of Next-Generation Sequencing Platforms. Pervez MT, Hasnain MJU, Abbas SH, Moustafa MF, Aslam N, Shah SSM. Biomed Res Int. 2022 Sep 29;2022:3457806. doi: 10.1155/2022/3457806. PMID: 36212714; PMCID: PMC9537002.

Individuals suffering from end-stage kidney disease (ESKD) undergoing dialysis have significantly higher mortality rates compared to patients with non-ESKD of the same age. Approaches to assess mortality risk, which could enable tailored and effective care are needed (1).

Angiopoietin-2 (Ang2) is a protein that is crucial in pathological vascular restructuring and in the formation of new blood vessels (angiogenesis). These processes are active in ESKD patients and might contribute to the elevated mortality observed in this patient group.

Angiopoietin-2 predicts mortality in male ESKD patients on hemodialysis

In a recent multicenter prospective study Ang2 was measured in 340 hemodialysis (HD) patients with ESKD to predict overall mortality (2, 3). The patients were followed up for 5-years and blood samples and clinical data were collected at baseline. Ang2 serum levels were measured with a enzyme-linked immunosorbent assay (ELISA) from Biomedica.

Angiopoietin-2 predicts mortality in male ESKD patients on hemodialysis

The results of the study demonstrated that the risk of death was higher in male patients with elevated Ang2 levels than in female patients: Ang2 levels at baseline are independently associated with all cause mortality in male ESKR patients on HD. 

Angiopoetin-2 can reliably be measured in serum or plasma samples with a conventional ELISA assay: 

Human ANGIOPOIETIN-2 ELISA (cat. no. BI-ANG2) 

• 20µl sample volume /well
• optimized for clinical samples – sample values provided
• kit includes ready to use standards and controls  
• developed & manufactured by BIOMEDICA
• for serum/plasma/cell culture samples
• validation package available

 

also available: 

Mouse / rat ANGIOPOIETIN-2 ELISA  (cat. no. BI-ANG2MR)

• 5µl sample volume /well
• kit control included
• sample values provided

 

Related Publications

1. One- and 2-Year Mortality Prediction for Patients Starting Chronic Dialysis. Haapio M, Helve J, Grönhagen-Riska C, Finne P. Kidney Int Rep. 2017 Jun 24;2(6):1176-1185. doi: 10.1016/j.ekir.2017.06.019. PMID: 29270526; PMCID: PMC5733880.
2. Angiopoietin-2 predicts all-cause mortality in male but not female end-stage kidney disease patients on hemodialysis. Chu C, Chen X, Hasan AA, Szakallova A, Krämer BK, Tepel M, Hocher B. Nephrol Dial Transplant. 2022 Jun 23;37(7):1348-1356. doi: 10.1093/ndt/gfab332. PMID: 34792167; PMCID: PMC9217660.
3. Circulating angiopoietin-2 level is independently associated with all-cause mortality in male end-stage kidney disease patients on hemodialysis. Chu, Chang. Thesis, 2023-06-22T11:52:24Z.

 

 

Lyme disease, also known as Lyme borreliosis, is a tick-borne infectious disease mainly caused by the bacterium species Borrelia burgdorferi.

The disease is primarily transmitted to humans through tick bites. Skin rash, fever, headache, fatigue, and joint pain are among the symptoms characterized by the disease. If left untreated, the infection can lead to more severe conditions and can spread to various parts of the body, affecting joint, heart, and nervous system. Lyme disease is most prevalent in temperate regions, particularly in North America, Europe, and parts of Asia. The incidence of Lyme disease has been increasing in recent years, making it an important health concern.

Early diagnosis and treatment are essential for managing Lyme disease effectively.

Testing for Lyme Borreliosis

The BIOMEDICA Borrelia ELISA kits were utilized in a recent study analyzing antibody patterns that could be useful in guiding the diagnostic schedule: Testing for Lyme borreliosis: could serology tell more? Zóka A et al., J Immunol Clin Microbiol. 2020; 5(3): 85-93. link .

Borrelia IgG ELISA | BI-21032

Recombinant Antigens– therefore:

• High sensitivity and specificity confirmed by clinical samples
• Standardized CSF and serum analysis available
• For manual and automated testing
• Widely cited

 

Lyme borreliosis is a bacterial infection caused by the spirochaete Borrelia burgdorferi, afzellii or garinii, and is characterised by a variety of clinical symptoms.

 Lyme borreliosis can be divided into 3 stages:

 Stage 1, early dermatitis, Clinical: erythema migrans.

Stage 2, early disseminated infection, Clinical: lymphocytic meningoradiculitis (Bannwarth’s syndrome), neuroborreliosis.

Stage 3, late disseminated infection, Clinical: chronic progressive encephalomyelitis, acrodermatitis chronica atrophicans (ACA), chronic arthritis.

To improve the diagnostic specifity, the Biomedica Borrelia IgG ELISA microwell strips are coated with the following recombinant antigens:

• p21 = OspC (outer surface protein C): B. burgdorferi sensu stricto (B31), B. garinii (20047)
• p18: B. afzelii (pKo)
• p100: B. afzelii (pKo)
• VIsE: fusion protein of different Borrelia genospecies

 

Borrelia IgM ELISA | BI-21042

Recombinant Antigens– therefore:

• No extra RF stripping necessary
• High sensitivity and specificity confirmed by clinical samples
• Standardized CSF-serum-analysis available
• For manual and automated testing
• Widely cited

 

To improve the diagnostic specifity, the Biomedica Borrelia IgM ELISA microwell strips are coated with the following recombinant antigens:

• p21 = OspC (outer surface protein C): B. afzellii (pKo)
• p21 = OspC (outer surface protein C): B. garinii (20047)
• p41/I = (inner part of flagellin): B. bavariensis (pBi)
• VIsE: fusion proteins of different Borrelia genospecies 

 

Related publications

Lyme borreliosis diagnosis: state of the art of improvements and innovations. Guérin M, Shawky M, Zedan A, Octave S, Avalle B, Maffucci I, Padiolleau-Lefèvre S. BMC Microbiol. 2023 Aug 1;23(1):204. doi: 10.1186/s12866-023-02935-5. PMID: 37528399; PMCID: PMC10392007.

The Epidemiology of Lyme Borreliosis in Europe: An Updated Review on a Growing Public Health Issue. Stark JH, Pilz A, Jodar L, Moïsi JC.Vector Borne Zoonotic Dis. 2023 Apr;23(4):139-141. doi: 10.1089/vbz.2022.0068. Epub 2023 Jan 27. PMID: 37071398; PMCID: PMC10122224.

Effectiveness of personal protection measures against Lyme disease: A review of epidemiologic studies from the United States. Schwartz AM, Mackeprang JM, Mead PS, Hinckley AF. Zoonoses Public Health. 2022 Nov;69(7):777-791. doi: 10.1111/zph.12984. Epub 2022 Jul 5. PMID: 35791092.

Breast cancer is the most common cancer in women worldwide. Approximately 15-20% of breast cancer cases are Human Epidermal Growth Factor Receptor (HER2)-positive. The overexpression of the HER2 protein promotes the growth and spread of cancer cells, indicating a poorer prognosis in these patients. Therapies, known as anti-HER2 drugs, specifically target the HER2 protein and inhibit its activity, thus improving the survival of patients with HER2 positive breast cancer. 

Evaluating Cardiotoxicity of HER-2 Targeted Drugs for Breast Cancer

One known side effect of HER-2 targeted therapies is cardiotoxicity which is characterized by left ventricular systolic dysfunction. Identifying cardiac dysfunction in HER2 positive breast cancer patients is crucial in order to prevent the development of heart failure in these patients. Monitoring cardiac function through imaging methods is mandatory for HER2-positive breast cancer patients. However, considering the possibility of cardiac damage even if there are no evident changes in heart function could be beneficial. Therefore, biomarkers could be useful in assessing early cardiac dysfunction in these patients.

Evaluating Cardiotoxicity of HER-2 Targeted Drugs for Breast Cancer: 

The cardiac biomarkers  NT-proBNP and NT-proANP

N-terminal pro-brain natriuretic peptide (NT-proBNP – amino acids 1-76) and N-terminal pro-atrial natriuretic peptide (NT-proANP – amino acids 1-98) are secreted from the cardiac ventricles and atria, respectively. The half-lives and stability of both analytes are more reliable than their biologically active peptides (BNP and ANP).

In a recent study researchers evaluated patients diagnosed with HER-2 positive breast cancer who underwent treatment with HER2 inhibitors and measured serum levels of the cardiac biomarkers NT-proBNP and NT-proANP using the assays from Biomedica. Learn more:

Evaluating Cardiotoxicity in Breast Cancer Patients Treated with HER2 Inhibitors: Could a Combination of Radionuclide Ventriculography and Cardiac Biomarkers Predict the Cardiac Impact?

Ready tp use kits developed & manufactured by BIOMEDICA

 

NT-proBNP ELISA Assay (cat. no SK-1204)

•  For the detection of human NT-proBNP
• CE.marked – for IVD use in the EU
• Flexible – can be run in every lab
• 20µl sample volume (serum/plasma)
• Assay time – 3 h / 30 min
• Two controls and ready to use calibrators included
• Regular proficiency testing –“Proficiency Testing Certificate”

 

NT-proANP ELISA Assay (cat. no BI-20892)

• For the detection of human NT-proANP (assay suitable also for detection of NT-proANP in rat and mouse samples)
• Two controls and ready to use calibrators included
• 50µl sample volume (serum/plasma)
• Assay time – 3 h / 30 min

 

Related products

Rat NT-proBNP ELISA assay (cat. no. BI-1204R)

• 10 µl sample volume
• sample values provided

 

Related literature

Evaluating Cardiotoxicity in Breast Cancer Patients Treated with HER2 Inhibitors: Could a Combination of Radionuclide Ventriculography and Cardiac Biomarkers Predict the Cardiac Impact? Gherghe M, Lazar AM, Mutuleanu MD, Bordea CI, Ionescu S, Mihaila RI, Petroiu C, Stanciu AE. Cancers (Basel). 2022 Dec 29;15(1):207. doi: 10.3390/cancers15010207. PMID: 36612202; PMCID: PMC9818586.

Abstract

(1) Background: The aim of our study was to determine whether monitoring cardiac function through RNV and cardiac biomarkers could predict the cardiac impact of combined therapy with trastuzumab, pertuzumab and docetaxel, which are regularly used nowadays to treat HER2-positive breast cancer. (2) Methods: This prospective monocentric study included 22 patients, diagnosed with HER2-positive breast cancer, who had their LVEFs and cardiac biomarkers evaluated both at the beginning of their treatment and after 6 months. Among all of the enrolled patients, two blood specimens were collected to assess circulating cardiac biomarkers. RNV was performed in each patient after “in vivo” radiolabeling of the erythrocytes. The obtained results were then statistically correlated. (3) Results: The average LVEF decrease between the two time points was approximately 4%. Of the five biomarkers we considered in this paper, only NT-proBNP correlated with the LVEF values obtained both in the baseline study and after 6 months of follow-up (r = -0.615 for T0 and r = -0.751 for T1, respectively). ST2/IL-33R proved statistically significant at the T1 time point (r = -0.547). (4) Conclusions: A combination of LVEF, NT-proBNP and ST2/IL-33R assessment may be useful for early detection of cardiac impairment in breast cancer patients treated with trastuzumab, pertuzumab and docetaxel.

NT-proBNP correlates with LVEF decline in HER2-positive breast cancer patients treated with trastuzumab. Bouwer NI, Liesting C, Kofflard MJM, Sprangers-van Campen SM, Brugts JJ, Kitzen JJEM, Fouraux MA, Levin MD, Boersma E. Cardiooncology. 2019 May 28;5:4. doi: 10.1186/s40959-019-0039-4. PMID: 32154011; PMCID: PMC7048136.

NT-proBNP as predictor factor of cardiotoxicity during trastuzumab treatment in breast cancer patients. Blancas I, Martín-Pérez FJ, Garrido JM, Rodríguez-Serrano F. Breast. 2020 Dec;54:106-113. doi: 10.1016/j.breast.2020.09.001. Epub 2020 Sep 11. PMID: 32977298; PMCID: PMC7511727.

We are scientists, developers and manufacturers and understand the importance of offering assays that generate specific, reliable and reproducible results.

Early on, starting from product development to final assay validation and product release until final ELISA kit manufacturing, every assay goes through a stringent quality control process.

Why choose ELISA assays from Biomedica?

Because we care!

Get the most accurate results from your precious samples with Biomedica ELISA kits.

   

 

SPECIFICIC – RELIABLE – REPRODUCIBLE ELISA Assays

 1. SPECIFICITY – epitope-mapped and characterized antibodies for accurate biomarker detection.

The performance of an ELISA is linked to the quality of the antibody pairs used for biomarker detection.

We therefore:

• select antibody pairs with high affinity and specificity with mapped binding sites
• optimize our assays to quantify biomarkers in both healthy and pathological samples

 

Example: FGF23 sample values of normal and pathological samples

2. RELIABILITY – extensive validation using clinical samples (parallelism, S/R, precision, analyte stability..) in various sample matrices

We validate our ELISA assays according to international quality guidelines (FDA, EMEA) .

Our Biomedica Immunassays go through an extensive validation process 

2.1. Accuracy – accurate detection of biomarkers in clinical samples and exclusion of matrix effects that may interfere with the measurement of the analyte of interest.

• Accuracy is determined in all validated sample types that 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.

 

2.2. Parallelism / Dilution Linearity – lot to lot consistency ensured by our stringent quality management guidelines

• During assay validation we analyze the recovery of the analyte in diluted samples that contains the endogenous / recombinant analyte of interest.

 

Example: dilution linearity (parallelism) of samples containing endogenous and recombinant Neuropilin-1 (NRP1).

2.3. Specificity and Cross-Reactivity – only detects the analyte of interest

• We carefully select antibodies that exclusively detect the specific analyte. Our specificity experiments are designed to characterize the antibody-antigen interactions and to determine possible isoforms that could be bound by the antibody.

 

Example: antibody recognizing all three isoforms of the target analyte on a Western blot.

 

2.4. Sensitivity

• Our ELISA assays are optimized to minimize the background signal while maximizing the signals from the measurements of the analyte ensuring maximal sensitivity.

The data for the Lower Limit of Quantification (LLOQ) and the Limit of Detection (LOD) are indicted in the instructions for use and on our website for all our ELISA kits.

2.5. Precision – precise and reproducible results within and across lots

• Within-run and in-between run precision is tested several times within one ELISA assay lot to guarantee that results are accurate when using kits that derive from different lots.

2.6. Calibration

• The accurate quantification of a biomarker depends on the linearity and the reproducibility of the standard curve. During the assays optimization process we ensure low variability between the results of the calibrators. Where available, we employ WHO reference reagents to ensure a harmonized standardization.

 

Example: standard curve for the FGF23 ELISA after 4PL transformation. The error bars reflect the variability of the measurement.

2.7. Stability

• During development we test the stability of all assay components as well as the stability of the analyte of interest in the respective sample matrices (serum, plasma). For instance we expose real clinical samples with elevated levels of the analyte to multiple freeze-thaw cyles and also determine stability at room temperature.

 

Example: stability of the analyte Periostin in clinical samples after multiple freeze-thaw (F/T) cycles in different sample matrices.

 

Freeze-thaw stability of Periostin

 

Validation reports

The validation reports of the respective ELISA assays can be  downloaded on the individual Biomedica ELISA product pages.

 

3. REPRODUCIBILITY

 ELISA ASSAY QUALITY MANAGEMENT

 Our Quality Control Process

All our kits undergo a stringent quality control process, including testing of lot-to-lot consistency as well as the kit stability during shelf-life.

Our manufacturing process follows the ISO 9001: 2015 management system and conforms to GMP /GLP guidelines.

Ensuring lot-to-lot consistency with a panel of quality control samples

Our internal quality control panel is one integral part of our manufacturing protocols. It contains samples from different matrices (serum, EDTA-plasma, citrate-plasma..) containing the endogenous/natural analyte as well as samples spiked with the recombinant protein. Every new lot as well as all retains, that are assayed every three months, are tested with the specific QC sample panel.

Example:  IC trending showing the quotient of the proANP concentration measured in Internal Controls (IC) in 3 different proANP ELISA lots compared with previously established median concentrations.

 

 

 

Qualified CUSTOMER SERVICE – we accompany you in every step.

Our qualified customer service representatives have hands-on research experience to assist you along the way, from decision making to technical questions.

WE VALUE YOUR OPINION

Our ELISA assays are developed to serve your needs. We therefore select our biomarker targets based on your input. 

Pre-testing: before a new ELISA assay is launched,  selected customers test the product to ensure that the assay is reliable and reproducible outside of our lab-facilities. 

Biomedica – ELISA development scheme  

 

Learn more about how we guarantee the performance of our products – click here .

 

Further reading

ICH Q2(R2) Validation of analytical procedures – Scientific guideline

 

Major depressive disorder (MDD) is a prevalent mental health condition that ranks among the top psychiatric illnesses worldwide. Approximately 30-60% of patients diagnosed with depression do not show positive responses to currently available antidepressant treatments. There is consistent evidence suggesting that elevated blood levels of the cytokine interleukin-6 (IL-6) in individuals with MDD have a significant impact on stress responses and correlate with depression severity scores (1, 2).

IL-6 in depressive disorder

Given the association between IL-6 and MDD, inhibiting IL-6 has gained attention as a potential therapeutic approach for depression. By targeting IL-6 pathways it is hypothesized that the aberrant inflammatory response associated with MDD can be modulated, leading to improved treatment outcomes. Exploring IL-6 inhibition, as a novel strategy holds promise for the development of more effective therapies for individuals who are non-responsive to current antidepressant treatments.

IL-6 can reliably be measured in various sample types by ELISA assay:

BIOMEDICA IL-6 ELISA (cat. no. BI-IL6)

• Sensitive – Measurable values in serum and plasma samples
• Reliable – Full validation package
• Specific-  Characterized epitope-mapped antibodies
• Easy – Color-coded reagents and controls included

 

Related ELISA products

VEGF (cat. no. BI-VEGF), Angiopoietin-2 (BI-ANG2), Big-Endothelin (cat. no. BI-20082H), NT-proCNP (cat. no. BI-20812)

Publications

1. How does IL-6 change after combined treatment in MDD patients? A systematic review. Lombardi AL, Manfredi L, Conversi D. Brain Behav Immun Health. 2022 Dec 24;27:100579. doi: 10.1016/j.bbih.2022.100579. PMID: 36624849; PMCID: PMC9822965.
2. Role of Interleukin-6 in Depressive Disorder. Ting EY et al., Int J Mol Sci. 2020 Mar 22;21(6):2194. doi: 10.3390/ijms21062194. PMID: 32235786; PMCID: PMC7139933.

 

Related literature

Interleukin 6 as a marker of depression in women with sleep apnea. Campos-Rodriguez F et al., J Sleep Res. 2021 Feb;30(1):e13035. doi: 10.1111/jsr.13035. Epub 2020 Mar 25. PMID: 32212220.

 

Abstract

Depression is common in women with obstructive sleep apnea (OSA), but objective markers of depression have not yet been explored in such patients. We hypothesized that inflammation and antioxidant biomarkers may be associated with depression in a cohort of OSA women. We conducted a multicentre, cross-sectional study in 247 women diagnosed with moderate-to-severe OSA. Depression was assessed by the depression subscale of the Hospital Anxiety and Depression Questionnaire (HAD-D) and defined as a score ≥11. Associations between tumour necrosis factor α (TNFα), interleukin 6 (IL-6), C-reactive protein (CRP), intercellular adhesion molecule 1 (ICAM-1), catalase (CAT), superoxide dismutase (SOD) and brain-derived neurotrophic factor (BDNF) plasma levels and depression were assessed. The women had a median (25th-75th percentiles) age of 58 (51-65) years, body mass index (BMI) of 33.5 (29.0-38.3) Kg/m² , Epworth Sleepiness Scale (ESS) score of 10 (6-13) and apnea-hypopnea index (AHI) of 33.3 (22.8-49.3). Logistic regression analyses revealed that only IL6 levels were associated with the presence of depression (adjusted odds ratio [OR], 1.20; 95% confidence interval [CI], 1.08-1.34), whereas linear regression further confirmed that IL6 levels were significantly associated with HAD-D scores (β = .154; 95% CI, 0.03-0.30). Multivariate regression analysis showed that IL6 (OR, 1.22; 95% CI, 1.09-1.36), ESS (OR, 1.10; 95% CI 1.02-1.19) and physical activity <30 min/day (OR, 2.51; 95% CI, 1.25-5.05) were independent predictors of depression. Thus, we conclude that in a cohort of women with moderate-to-severe OSA, IL6 levels are independently associated with the presence of depression and correlate with depression scores. Low physical activity and higher ESS scores are also independent indicators of risk of depression in this population.

The process of aging is linked to physiological changes, which include a decline in bone mass and renal function. In a recent study, researchers investigated the effects of Vitamin D supplementation on bone metabolism and bone biomarkers in patients with and without renal impairment.

Effects of Vitamin D on Bone Markers in Kidney Disease

The study incorporated 379 patients with a mean age of > 70 years who were supplemented with various doses of vitamin D ranging from 12000 IU to 48,000 IU/month for a period of one year.

The biomarkers Sclerostin (SOST), Dickkopf-1 (DKK-1), Osteoprotegerin (OPG), and soluble RANKL (sRANKL) were measured with ELISA assays from BIOMEDICA.

Learn more: Early renal impairment affects hormonal regulators of calcium and bone metabolism and Wnt signalling and the response to vitamin D supplementation in healthy older adults. Christodoulou M et al., J Steroid Biochem Mol Biol. 2023. 229:106267. doi: 10.1016/j.jsbmb.2023.106267. 

About the BIOMEDICA bone marker ELISA kits

Sclerostin (SOST) ELISA  (BI-20492)

• Most referenced Sclerostin ELISA + 250 citations
• Low sample volume – 20µl / well
• Validated following international guidelines

 

Dickkopf-1 (DKK-1) ELISA  (BI-20413)

• Widely cited + 170 publications
• Direct measurement
• Validated following international guidelines

 

Osteoprotegerin (OPG) ELISA (BI-20403)

• most referenced human OPG ELISA in +245 citations
• day test, ready to use color coded reagents
• controls included

 

Free soluble RANKL ELISA (BI-20462)

• Highly sensitive  – measurable concentrations in healthy subjects
• Only ELISA that measures free, uncomplexed soluble RANKL
• Cited in over + 290 publications

 

FGF23 intact ELISA (BI-20700)

• for serum and plasma samples
• full validation data available
• citations

 

FGF23 c-terminal ELISA (BI-20702)

• for serum and plasma
• full validation data available
• + 43 citations

 

All assays are developed and manufactured by BIOMEDICA

Related publications

Fibroblast growth factor 23 (FGF23) and early chronic kidney disease in the elderly. Chudek J et al., Nephrol Dial Transplant. 2014 Sep;29(9):1757-63. doi: 10.1093/ndt/gfu063. 

Effectiveness and safety of vitamin D in relation to bone health. Cranney A et al., Evid Rep Technol Assess (Full Rep). 2007 Aug;(158):1-235. PMID: 18088161; PMCID: PMC4781354.

Bone markers are currently used to monitor skeletal diseases and treatments. The proteins Sclerostin and Dickkopf-1 (DKK-1) reflect distinct biological processes and have gained attention as potential biomarkers for bone-related conditions. They may provide valuable information for diagnosis, prognosis, and monitoring of bone diseases and treatments.

Sclerostin and DKK-1 emerging biomarkers for bone disease

Sclerostin and Dickkkopf-1 are two important osteocyte proteins that are involved in the regulation of bone metabolism, particularly through their interactions with the Wnt signaling pathway.

SCLEROSTIN (SOST) is a glycoprotein that is primarily secreted by osteocytes, the most abundant cells in bone tissue. It inhibits Wnt signaling, which is a critical pathway regulating bone formation and remodeling. Sclerostin acts as a negative regulator of bone formation by binding to the LRP5/6 co-receptors (low-density lipoprotein receptor protein), which activate Wnt signaling. By binding to LRP5/6, sclerostin inhibits the interaction between Wnt ligands and the Frizzled receptor, thereby inhibiting Wnt signaling and suppressing bone formation. Inhibition of Sclerostin has led to the development of a novel anabolic therapy for osteoporosis.

DICKKOPF-1 (DKK-1) is a protein that also inhibits the Wnt signaling pathway. DKK-1 binds to the LRP5/6 co-receptors thereby preventing Wnt ligand interaction thus inhibiting bone formation and promoting bone resorption.

The Wnt-signaling pathway is one of the most important pathways controlling bone metabolism. Sclerostin and Dickkopf-1 act as Wnt inhibitors and play a crucial role in controlling bone formation and resorption.

Sclerostin and DKK-1 can easily be measured in blood samples with an ELISA assay

BIOMEDICA offers two kits to measure Sclerostin

Bioactive Sclerostin ELISA cat. no. BI-20472

• serum, plasma, cell-culture
• characterized antibodies
• day test
• 20µl sample volume
• sample values provided
• full validation package

 

The epitope of the monoclonal capture antibody is in loop 2 of the Sclerostin molecule, which is the binding site for the LRP5/6 complex.

All Sclerostin molecules including potential fragments containing this receptor binding region can be detected.

Bioactive Sclerostin ELISA – antibody binding sites

 

Sclerostin ELISA cat. no. BI-20492

• most referenced more than 280 citations
• 20µl sample volume
• for serum, plasma
• full validation package

 

DKK-1 ELISA cat. no. BI-20413

• widely cited
• day test
• no sample dilution
• full validation package

 

All assays are developed and manufactured by BIOMEDICA

Related Publications

New Emerging Biomarkers for Bone Disease: Sclerostin and Dickkopf-1 (DKK1). Dincel AS, Jørgensen NR; IOF-IFCC Joint Committee on Bone Metabolism (C-BM). Calcif Tissue Int. 2023 Feb;112(2):243-257. doi: 10.1007/s00223-022-01020-9. Epub 2022 Sep 27. PMID: 36165920.

Abstract

A healthy skeleton depends on a continuous renewal and maintenance of the bone tissue. The process of bone remodeling is highly controlled and consists of a fine-tuned balance between bone formation and bone resorption. Biochemical markers of bone turnover are already in use for monitoring diseases and treatment involving the skeletal system, but novel biomarkers reflecting specific biological processes in bone and interacting tissues may prove useful for diagnostic, prognostic, and monitoring purposes. The Wnt-signaling pathway is one of the most important pathways controlling bone metabolism and consequently the action of inhibitors of the pathway such as sclerostin and Dickkopf-related protein 1 (DKK1) have crucial roles in controlling bone formation and resorption. Thus, they might be potential markers for clinical use as they reflect a number of physiological and pathophysiological events in bone and in the cross-talk with other tissues in the human body. This review focuses on the clinical utility of measurements of circulating sclerostin and DKK1 levels based on preanalytical and analytical considerations and on evidence obtained from published clinical studies. While accumulating evidence points to clear associations with a number of disease states for the two markers, and thus, the potential for especially sclerostin as a biochemical marker that may be used clinically, the lack of standardization or harmonization of the assays still hampers the clinical utility of the markers.

 

Therapeutic approaches to activate the canonical Wnt pathway for bone regeneration. Nelson AL, Fontana G, Miclau E, Rongstad M, Murphy W, Huard J, Ehrhart N, Bahney C. J Tissue Eng Regen Med. 2022 Nov;16(11):961-976. doi: 10.1002/term.3349. Epub 2022 Sep 16. PMID: 36112528; PMCID: PMC9826348.

The BIOMEDICA FGF23 ELISA assays were recently used in a study investigating hepcidin and iron status in patients with inflammatory bowel disease undergoing therapy (1). Hepcidin, a peptide produced by liver cells, plays a vital role in regulating the body´s iron levels. In instances of inflammation, circulating hepcidin levels rise. Inflammation and iron deficiency have also been shown to stimulate FGF23 production (2). Iron deficiency enhances both the transcription and post-translational cleavage of FGF23 (3). Typically, this results in increased serum FGF23 C-terminal levels, with little to no effect on the biologically active intact FGF23 concentrations (4).

About FGF23

Fibroblast growth factor 23 (FGF23) is a protein that plays a crucial role in regulating phosphate and vitamin D metabolism in the body.

It is primarily produced by bone cells in the tissue called osteocytes. FGF 23 acts on the kidneys and decreases phosphate reabsorption thereby preventing the excessive accumulation of phosphate in the body.

Inflammatory bowel disease and FGF23

Patients with inflammatory bowel disease (IBD) face a greater risk to develop osteopenia and osteoporosis compared to the general population (5). In a study investigating the role of FGF23 in childhood inflammatory bowel disease, serum FGF23 was significantly higher in patients with IBD compared to controls (6)

How can you quantify circulating FGF23 levels?

FGF23 can easily be measured in blood samples (serum or plasma) with a conventional ELISA assay. The levels of FGF23 in the bloodstream consist of both the active intact hormone (iFGF23) and the inactive c-terminal fragments (cFGF23).

The C-terminal FGF23 assay captures both the intact FGF23 hormone and the fragments that form after FGF23 has been cleaved. The intact FGF23 assays utilize antibodies that are positioned in the N-terminal and C-terminal regions of the hormone, specifically targeting the biologically active intact FGF23.

Read more :  FGF23 an Overview

BIOMEDICA  FGF23 ELISA kits for  serum & plasma samples

FGF23 intact ELISA, # BI-20700

FGF23 (c-terminal) ELISA, # BI-20702

• developed & manufactured by Biomedica , Austria
• fully validated according to international quality guidelines
• numerous top journal citations

 

Related products : INTERLEUKIN-6  (IL-6) ELISA , #BI-IL6

• highly sensitive-measurable values in both serum and plasma
• ready to use calibrators and controls included

 

Literature

1. Hepcidin and Iron Status in Patients With Inflammatory Bowel Disease Undergoing Induction Therapy With Vedolizumab or Infliximab. Loveikyte R, Bourgonje AR, van der Reijden JJ, Bulthuis MLC, Hawinkels LJAC, Visschedijk MC, Festen EAM, van Dullemen HM, Weersma RK, van Goor H, van der Meulen-de Jong AE, Dijkstra G. Inflamm Bowel Dis. 2023 Feb 7:izad010. doi: 10.1093/ibd/izad010. Epub ahead of print. PMID: 36748574.
2. Inflammation and functional iron deficiency regulate fibroblast growth factor 23 production. David V, Martin A, Isakova T, Spaulding C, Qi L, Ramirez V, Zumbrennen-Bullough KB, Sun CC, Lin HY, Babitt JL, Wolf M. Kidney Int. 2016 Jan;89(1):135-46. doi: 10.1038/ki.2015.290. Epub 2016 Jan 4. PMID: 26535997; PMCID: PMC4854810.
3. Regulation of Fibroblast Growth Factor 23 by Iron, EPO, and HIF. Wheeler JA, Clinkenbeard EL. Curr Mol Biol Rep. 2019 Mar;5(1):8-17. doi: 10.1007/s40610-019-0110-9. Epub 2019 Jan 25. PMID: 31218207; PMCID: PMC6582956.
4. Regulation of FGF23: Beyond Bone. Simic P, Babitt JL.Curr Osteoporos Rep. 2021 Dec;19(6):563-573. doi: 10.1007/s11914-021-00703-w. Epub 2021 Nov 10. PMID: 34757587; PMCID: PMC8958553.
5. Advances in the understanding of mineral and bone metabolism in inflammatory bowel diseases. Ghishan FK, Kiela PR. Am J Physiol Gastrointest Liver Physiol. 2011 Feb;300(2):G191-201. doi: 10.1152/ajpgi.00496.2010. Epub 2010 Nov 18. PMID: 21088237; PMCID: PMC3043650.
6. Fibroblast growth factor 23 contributes to diminished bone mineral density in childhood inflammatory bowel disease. El-Hodhod MA, Hamdy AM, Abbas AA, Moftah SG, Ramadan AA. BMC Gastroenterol. 2012 May 2;12:44. doi: 10.1186/1471-230X-12-44. PMID: 22551310; PMCID: PMC3438067.

Rat NT-proBNP and NT-proANP ELISA assays for drug discovery and translational research

Cardiac toxicity is a leading cause of preclinical safety failures in drug development. The cardiac markers NT-proANP and NT-proBNP have proven to be useful in preclinical toxicology testing.

Cardiac Safety Biomarker Assays in Preclinical Toxicology Testing

 

The BIOMEDICA NT-proANP and NT-proBNP ELISA kits are robust assays to quantify these cardiac hormones in rat samples. Due to the high inter-species homology the NT-proANP kit has also been applied in canine and feline samples.

BIOMEDICA´s NT-proANP ELISA kit (BI-20892) is widely published and has been independently validated for cardiovascular safety studies in rats (1, 2).

Rat NT-proBNP ELISA assay (cat. no. BI-1204R) NEW

contact us for your special evaluation discount  info@bmgrp.com

• 10 µl / well,  serum or plasma
• kit control included
• sample values provided

 

NT-proANP ELISA assay (cat. no. BI-20892)  

• 10 µl / well, serum or plasma
• widely cited as cardiovascular safety biomarker in rats
• for use in human and non-human samples (high cross-reactivity between species)

 

PRODUCT INFORMATION

Rat NT-proBNP ELISA  

• Product code: BI-1204R
• Method: ELISA
• Time to result: 3.5 hours
• Sample types: rat serum and plasma
• Sample volume: 10 µl/well
• Sensitivity LOD: 21 pg/ml
• Standard curve range: 0 – 3200 pg/ml
• Specificity: Endogenous and recombinant rat NT-proBNP
• Instructions for use click here

 

NT-proANP ELISA 

• Product code: BI-20892
• Method: ELISA
• Time to result: 3.5 hours
• Sample types: Serum, plasma, urine, cell culture supernatant (human, rat, mouse 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 and recombinant human NT-proANP (equivalent to proANP 1-98). Very high sequence homology between human and rodent (rat, mouse, and other species e.g. rabbit).
• NT-proANP ELISA assay is suitable for rat and mouse samples
• Instructions for use: click here

 

CARDIAC SAFETY BIOMARKER ASSAYS in PRECLINICAL TOXICOLOGY TESTING 

References/Citations/Related Literature

NT-proANP as a cardiovascular safety biomarker in rat and mouse samples , click for all references

1. 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 Jan-Feb;77:58-65. doi: 10.1016/j.vascn.2015.10.002. Epub 2015 Oct 26. PMID: 26516096.
2. Natriuretic Peptides as Cardiovascular Safety Biomarkers in Rats: Comparison With Blood Pressure, Heart Rate, and Heart Weight. Engle SK, Watson DE.Toxicol Sci. 2016 Feb;149(2):458-72. doi: 10.1093/toxsci/kfv240. Epub 2015 Nov 25. PMID: 26609138.

 

Abstract

When using an ELISA assay, it is important to ensure that the results are specific, accurate, sensitive, and reproducible. The ELISA assay reliability  varies among kit suppliers, so choosing the “right” ELISA requires careful consideration. Access to the assay´s validation data before making a choice may be helpful.

ELISA Assay Reliability

At BIOMEDICA we take pride in developing and manufacturing our ELISA kits following a stringent manufacturing and quality control process, that enables us to maintain consistent and reproducible outcomes with every lot we produce.

At Biomedica we develop and manufacture our ELISA assays with care

 

All BIOMEDICA kits are fully validated and come with sample data for healthy human subjects, ready-to-use standards and controls.  We supply our kits with color-coded vials, and some of the reagents are  “colorful” as well, to make the kits easy to use and to avoid possible pipetting errors, To increase transparency, we publish the validation data of every ELISA assay kit on our website. 

BIOMEDICA´s  QUALITY GUIDELINES

AT BIOMEDICA, our goal is to supply reliable and well-validated ELISA kits for your research.

Here is how we ensure product excellence:

1. Optimization: we diligently optimize all Biomedica assay to guarantee reliability, sensitivity, and precision.
2. Validation: our kits undergo an extensive validation process in accordance with international quality guidelines (FDA, EMA, and ICH), ensuring the kits accuracy and efficiency.
3. Expert Team: our dedicated team consists of highly qualified scientists, many with doctorate-level education and industry training. Our team has extensive research experience and contributes to the development, production, and customer service aspects of our products.
4. Quality Management: Biomedica adheres to the ISO 9001: 2015 certified quality management system in our manufacturing process, ensuring consistent and high-quality products.

 

With these measures in place, we are committed to deliver ELISA assays that meet the highest standards of performance and reliability.

Learn more: https://www.bmgrp.com/quality 

Related literature

Interference in ELISA. Matson RS. 2023. Methods Mol Biol. 2612:91-99. doi: 10.1007/978-1-0716-2903-1_7. PMID: 36795361.

Abstract

Practical Guide to Immunoassay Method Validation. Andreasson U et al. 2015. Front Neurol. 19;6:179. doi: 10.3389/fneur.2015.00179. PMID: 26347708; PMCID: PMC4541289.

 

EZ4U –  Easy and Reliable Assay for Cell Viability Analysis

Cell viability quantifies the percentage of living and healthy cells within a given cell population. Cell viability analysis assays are employed to assess cell survival for instance after treatment with substances during drug screening.

Testing of Cell Viability with Biomedica´s EZ4U Assay

The EZ4U assay is a metabolic assay (like the MTT assay) that quantifies cell health by measuring the reduction of the colorimetric substrate through the activity of mitochondrial enzymes.

Cell viability assays are widely used in cell biology research. The EZ4U test kit is a reliable and straightforward non-radioactive assay that can be completed within two to five hours, depending on the cell type studied. The EZ4U assay employs non-toxic tetrazolium salts, which are 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. Furthermore, as the assay procedure is identical to thymidine incorporation procedure, no changes in test protocols are necessary. 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. For examples please follow the citations by clicking this link.

EZ4U Cell Proliferation and  Cytotoxicity Assay –straightforward with a single incubation step using living cells

The EZ4U assay (Biomedica, Austria) was used in a recent study investigating the effects of RNA methylation, namely m⁶A methylation, on the renal cell carcinoma (ACHN and 769P) cell lines. Renal cell carcinoma (RCC) accounts for about 2% of cancer-related deaths. Patients with metastatic RCC have a poor prognosis with an approximate 5% survival rate. Despite therapeutical advancements, including checkpoint immunotherapy,  the improvement in patient survival rates has been rather moderate.

Learn more: 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, Klümper N, Schmidt D, Ritter M, Ellinger J, Hauser S. Am J Transl Res. 2023 Mar 15;15(3):1744-1755. PMID: 37056835; PMCID: PMC10086911.

Abstract

Background: N⁶-methyladenosine (m⁶A) is one of the most common RNA modifications in eukaryotes and has effects on RNA structure and stability. Recent studies have shown that m⁶A methylation is involved in human carcinogenesis. In the present study, we investigated the effects of m⁶A demethylases FTO and ALKBH5 on renal cell carcinoma (RCC) cell lines. Methods: The epithelial-mesenchymal in vitro knockdowns of FTO and ALKBH5 induced by antisense oligonucleotides (LNA-GapmeR system) were established in RCC cell lines. Their effects on migration and proliferation were investigated subsequently. The influence of FTO and ALKBH5 knockdown on key epithelial-mesenchymal transition (EMT) genes was analyzed. Results: Inactivation of FTO and ALKBH5 resulted in decreased proliferation and motility in all cell lines examined (ACHN, Caki-1, 769-P). Vimentin (VIM) was downregulated after the knockdown of FTO and ALKBH5, indicating an EMT switch. Conclusions: Knockdown of the m⁶A erasers FTO and ALKBH5 inhibits the malignant potential in the cell cultures studied by means of an EMT switch.

EZ4U  Cell Proliferation & Cytotoxicity Assay (cat.no. BI-5000)

 

• Non-radioactive & non-toxic assay
• Reliable & Sensitive
• Convenient single-step incubation  – for use on living cells
• Widely cited in more than 240 publications

 

Find out more: BROCHURE – EZ4U cell proliferation and cytotoxicity assay

 

 

 

 

At Biomedica we develop and manufacture quality ELISAs to quantify cytokine release

Our ELISAs to quantify cytokine release are validated according to international quality guidelines to ensure their consistency, specificity, precision, and robustness. The validation data files can be found on the respective product pages www.bmgrp.com. 

Our kits are designed to specifically quantify cytokine release in various biological matrices (e.g. serum, plasma, cell culture).

WHO reference reagents for a harmonized standardization

All our cytokine kits are standardized using WHO reference reagents/standards to allow cross-comparison of results when using different reagent sets during a study (1).

BIOMEDICAs CYTOKINE ELISA KITS

 

IL-6 ELISA (cat. no. BI-IL6)

The Biomedica IL-6 (interleukin-6) ELISA kit is a sandwich ELISA that incorporates two epitope-mapped antibodies that specifically bind to human IL-6 in the respective samples.

Capture antibody (pre-coated on a 96-well microtiter plate): recombinant IL-6 antibody (specific for human IL-6).

Detection antibody: polyclonal IL-6 antibody (streptavidin-HRPO labeled), specific for human IL-6.

• • Method: sandwich ELISA
  • Sample type: serum, plasma, cell-culture, urine
  • Sample volume: 100µl / well
  • Assay time: 4.5 hrs
  • Standard range: 0 / 3.1 – 200 pg/ml
  • Sensitivity: LOD: 0.28 pg/ml; LLOQ: 0.78 pg/ml (measurable concentrations in serum AND plasma samples!)
  • Precision: in-between-run (n=9 ): ≤ 6 % CV, within-run (n=3): ≤ 7 % CV
  • Specificity:  the ELISA recognizes recombinant and endogenous (natural) human IL-6
  • Standard matrix: serum based matrix containing recombinant IL-6 including 8x vials of pre-diluted standards, lyophilized
  • Controls: 2 controls (high and low) included

 

VEGF ELISA (cat. no. BI-VEGF)

The Biomedica VEGF (Vascular Endothelial Growth Factor) ELISA kit is a sandwich ELISA that incorporates two epitope-mapped antibodies that specifically bind to human VEGF in the respective samples.

Capture antibody (pre-coated on a 96-well microtiter plate): recombinant VEGF antibody (specific for human VEGF) recognizes a structural epitope in the conserved receptor binding- site of the VEGF peptide. The antibody binds to all VEGF isoforms.

Detection antibody: polyclonal VEGF antibody (streptavidin-HRPO labeled), specific for human VEGF, recognizing multiple linear epitopes that are concentrated in the first 120 amino acids in the N-terminal region of the VEGF molecule.   

• Method: sandwich ELISA
• Sample type: serum, plasma, cell-culture, urine
• Sample volume: 10µl / well
• Assay time: 4.5 hrs
• Standard range: 0 / 31.2 – 2000 pg/ml
• Sensitivity: LOD: 2.5 pg/ml; LLOQ: 15.6 pg/ml (measurable concentrations in serum AND plasma samples)
• Precision: in-between-run: (n=3): ≤ 6 % CV, Within-run (n=3): ≤ 3 % CV
• Specificity: the ELISA recognizes recombinant and endogenous (natural) human VEGF including all circulating VEGF isoforms including VEGF165b
• Standard matrix: Serum based matrix containing recombinant VEGF, including 8x vials of pre-diluted standards, lyophilized
• Controls: 2 controls (high and low) included

 

ANGIOPOIETIN-2 ELISA (cat. no. BI-ANG2)

The Biomedica ANG2 (Angiopoietin-2) ELISA kit is a sandwich ELISA that incorporates two epitope-mapped antibodies that specifically bind to human Angiopoietin-2 in the respective samples.

Capture antibody (pre-coated on a 96-well microtiter plate): recombinant ANG2 antibody (specific for human ANG2).The capture antibody has a structural epitope that binds to the receptor binding site of human Angiopoietin-2. The receptor binds to TEL / TIE2 protein.

Detection antibody: polyclonal ANG2 antibody (streptavidin-HRPO labeled), specific for human ANG2, recognizing several linear epitopes that are distributed over the entire Angiopoietin-2 molecule.  

• Method: sandwich ELISA
• Sample type: serum, plasma, cell-culture, urine
• Sample volume: 20µl / well
• Assay time: 3.5 hrs
• Standard range: 0 / 12.5 – 400 pg/ml
• Sensitivity: 3.7 pmol/l (= 203 pg/ml)
• Precision: In-between-run (n=9): ≤ 6 % CV, Within-run (n=3): ≤ 8 % CV
• Specificity: the ELISA recognizes recombinant and endogenous (natural) human ANG2.  The assay most probably detects all three ANG2 isoforms, as determined by epitope mapping and analysis of the ANG2 sequence. ANG1 is not detected in this ELISA assay.
• Standard matrix: Serum based matrix containing recombinant ANG2,  including 8x vials of pre-diluted standards, lyophilized
• Controls: 2 controls (high and low) included

 

Also available: Mouse /rat Angiopoietin-2 ELISA (cat. no. BI-ANG2MR)

 

Related publications

1.       International reference reagents 

2.       Targeting IL-6 trans-signalling: past, present and future prospects. Rose-John S, Jenkins BJ, Garbers C, Moll JM, Scheller J. Nat Rev Immunol. 2023 Apr 17:1–16. doi: 10.1038/s41577-023-00856-y. Epub ahead of print. PMID: 37069261; PMCID: PMC10108826.

Abstract

Interleukin-6 (IL-6) is a key immunomodulatory cytokine that affects the pathogenesis of diverse diseases, including autoimmune diseases, chronic inflammatory conditions and cancer. Classical IL-6 signalling involves the binding of IL-6 to the membrane-bound IL-6 receptor α-subunit (hereafter termed ‘mIL-6R’) and glycoprotein 130 (gp130) signal-transducing subunit. By contrast, in IL-6 trans-signalling, complexes of IL-6 and the soluble form of IL-6 receptor (sIL-6R) signal via membrane-bound gp130. A third mode of IL-6 signalling – known as cluster signalling – involves preformed complexes of membrane-bound IL-6-mIL-6R on one cell activating gp130 subunits on target cells. Antibodies and small molecules have been developed that block all three forms of IL-6 signalling, but in the past decade, IL-6 trans-signalling has emerged as the predominant pathway by which IL-6 promotes disease pathogenesis. The first selective inhibitor of IL-6 trans-signalling, sgp130, has shown therapeutic potential in various preclinical models of disease and olamkicept, a sgp130Fc variant, had promising results in phase II clinical studies for inflammatory bowel disease. Technological developments have already led to next-generation sgp130 variants with increased affinity and selectivity towards IL-6 trans-signalling, along with indirect strategies to block IL-6 trans-signalling. Here, we summarize our current understanding of the biological outcomes of IL-6-mediated signalling and the potential for targeting this pathway in the clinic.

 

Fibroblast growth factor 23 (FGF23) is a hormone that plays a crucial role in regulating serum phosphate and vitamin D levels within the body. FGF23 is primarily produced in the bone by osteocytes and osteoblasts in response to factors such as oral phosphate intake or elevated serum Vitamin D concentrations. The regulation of normal serum phosphorus levels is primarily achieved through a highly controlled process occurring in the kidney where phosphate reabsorption takes place. Patients suffering from chronic kidney disease (CKD) have high plasma FGF23 levels and FGF23 serves as a sensitive biomarker for detecting abnormal renal phosphate handling. Notably, FGF23 levels increase during the early stages of kidney dysfunction, providing valuable insights into the underlying renal impairments (1, 2).

Effects of FGF23 on the heart

High FGF23 concentrations have been found to be associated with multiple cardiac diseases, including left ventricular hypertrophy, heart attacks, heart failure, and cardiovascular related deaths (3-5).

Present findings exploring the relationship of FGF23 and cardiac events are presented in a recent review. The authors further discuss the potential mechanisms by which FGF23 directly or indirectly triggers left ventricular hypertrophy (6). 

Learn more: Direct and indirect effects of fibroblast growth factor 23 on the heart. Nakano T et al., 2023.

Download our leaflet “FGF23 – an overview” here

How can FGF23 be measured?

Circulating FGF23 concentrations can be measured through blood tests by immunoassays. A widely used method for measuring FGF23 is with an ELISA assay (enzyme-linked immunosorbent assays). FGF23 ELISA assays utilize specific antibodies that recognize and bind FGF23 present in the sample.

Currently, there are two different assays commercially available for measuring circulating FGF23 in blood samples (7).

Intact FGF23 Assays

intact FGF23 (iFGF23) represents the full length, biologically active form of the hormone. It consists of the complete FGF23 protein structure without being enzymatically cleaved. The two antibodies in these assays target the N-terminal part and the C-terminal domain of the FGF23 molecule, respectively.

FGF23 C-terminal Assays

The c-terminal fragments of FGF23 (cFGF23) are the result of the enzymatic cleavage of the intact FGF23 molecule. The antibodies utilized in the cFGF23 assays bind to specific epitopes that are located in the c-terminal domain of the FGF23 molecule.

Noteworthy, all commercially available FGF23 c-terminal assays detect both c-terminal FGF23 fragments as well as the intact FGF23 molecule (7).

BIOMEDICA has developed two distinct ELISA assays to reliably quantify FGF23 concentrations in human serum and plasma.

·       FGF23 intact ELISA (cat. no. BI-20700)

·       FGF23 (C-terminal) ELISA (cat. no. BI-20702)

• Validated FGF23 ELISA kits according to international quality guidelines
• Numerous references in top-ranking journals

 

All Assays are Developed & Manufactured by Biomedica

Literature

1. The regulation of FGF23 under physiological and pathophysiological conditions. Rausch S, Föller M. Pflugers Arch. 2022 Mar;474(3):281-292. doi: 10.1007/s00424-022-02668-w. Epub 2022 Jan 27. PMID: 35084563; PMCID: PMC8837506.
2. Regulation and Effects of FGF23 in Chronic Kidney Disease. Musgrove J, Wolf M. Annu Rev Physiol. 2020 Feb 10;82:365-390. doi: 10.1146/annurev-physiol-021119-034650. Epub 2019 Nov 19. PMID: 31743079.
3. FGF23 and Cardiovascular Risk. Prié D. Ann Endocrinol (Paris). 2021 Jun;82(3-4):141-143. doi: 10.1016/j.ando.2020.03.007. Epub 2020 Mar 10. PMID: 32950228.
4. FGF23 predicts outcomes in heart failure but questions remain unanswered. Duran A, daSilva-deAbreu A, Joury A, Ventura HO. Int J Cardiol. 2021 Sep 1;338:145-146. doi: 10.1016/j.ijcard.2021.06.036. Epub 2021 Jun 19. PMID: 34157358.
5. Paracrine Effects of FGF23 on the Heart. Front Endocrinol (Lausanne). Leifheit-Nestler M, Haffner D 2018 May 28;9:278. doi: 10.3389/fendo.2018.00278. PMID: 29892269; PMCID: PMC5985311.
6. 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.
7. The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Calcif Tissue Int. Heijboer AC, Cavalier E. 2023 Feb;112(2):258-270. doi: 10.1007/s00223-022-00987-9. Epub 2022 Jun 4. PMID: 35665817; PMCID: PMC9859838.

and become a product reviewer

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We would love to hear from you! Start your product review here:  https://buff.ly/2UbmvFU

By the way, every reviewer receives an Amazon Gift Card.  

Check out some of the Biomedica product reviews.  Your review is highly appreciated – a big thank you!

• Reliable Assay Kit
• Easy, Reliable NT-ProANP ELISA Kit
• Easy-to-Use System that Delivers the Desired Results
• Measuring Cell Proliferation with EZ4U

 

 

YOUR SATISFACTION IS OUR PRIORITY

Our goal is to meet your needs at every stage, starting from assay design and development, through manufacturing, validation and customer support. We have implemented a range of measures to achieve this:

• Ready-to-Use Assays
• Biomarker Detection in Real Samples
• Validation According to International Standards
• Qualified, Experienced Customer Service
• Flexibility
• Service Measurements

 

learn more “About Biomedica” 

QUALITY is not just a word

Rest assured when using our products – we prioritize quality throughout the entire process from product development to manufacturing and quality control. Our goal is to ensure that the performance of our products meet the top quality standards.

Right after product development our kits go through a rigorous validation process in compliance with the international quality guidelines (FDA, EMEA, ICH).  Each product is accompanied by a comprehensive validation report that can downloaded from our website www.bmgrp.com .

WE ARE HERE FOR YOU

The vast majority of Biomedica products is delivered and used without any issues. However, if you do encounter a problem or have a question, we are here to help. Our dedicated and qualified team will do their best to answer your questions and will be there for you. Due to the close collaboration between our team members in development and production, we are in a unique position to support our customers with all of the detailed information they may need.

Being scientists at heart, we are always happy to share our knowledge, give advice, and help our customers selecting biomarkers.

 

Role of C-Type Natriuretic Peptide (CNP) in bone growth and development

CNP is a member of the natriuretic peptide family. It acts as a paracrine hormone which is present in several tissues such as the cardiovascular system, brain and the endothelium. CNP controls various processes including blood pressure, water and mineral balance, and numerous metabolic functions (1). In 1995, CNP was first described as a potent regulator of bone remodeling, underlying its important role in humans (2).

C-Type Natriuretic Peptide Induces Bone Growth

Research has indicated that CNP is predominantly synthesized within the joint cartilage by chondrocytes, which are specialized cells primarily responsible for generating collagen and the extracellular matrix. As a result, the primary and well-established role of CNP is in the growth plate skeletal cartilage, where it plays a crucial role in promoting endochondral growth of the developing skeleton (3, 4).

What is the mechanism of bone growth?

The growth of bones is a complex process involving the formation and remodeling of bone tissue throughout the body. It is regulated by a diverse range of hormones, growth factors, and signaling pathways that interact together to maintain skeletal health and function.  

Bone growth can be broadly classified into two types:

Longitudinal growth primarily takes place in the long bones of the body, such as femur and tibia. It is a process by which bones grow in length, predominantly occurring during childhood and adolescence. Longitudinal growth is facilitated by growth plates, which are cartilaginous areas located at the ends of long bones. As the bones elongate, the cartilage in the growth plates gradually undergoes ossification, transforming into bone tissue.

 Appositional growth, on the other hand, is the process by which bones grow in thickness or diameter. This type of growth occurs throughout life and is regulated by a delicate balance between osteoblasts, cells responsible for bone formation and osteoblasts, cells involved in bone resorption and bone-resorbing cells named osteoclasts.

Numerous factors can influence bone growth, including genetics, nutrition, exercise, and hormonal equilibrium. Hormones such as growth hormone and sex hormones play crucial roles in regulating bone growth and development. One hormone that is of particular significance in this regard is C-type natriuretic peptide (CNP), which is essential for the regulation of bone growth.

Quantification of CNP and NT-proCNP in human blood samples

The measurement of biologically active CNP in human blood samples presents a challenge due to its dynamic role in bone growth, its rapid clearance and low circulating concentrations. However, the inactive, but more stable precursor molecule NT-proCNP, has proven to be a reliable alternative. The amino-terminal propeptide NT-proCNP, is synthesized in the growth plate, has a longer half-life and circulates in equimolar concentrations as CNP. Therefore, NT-proCNP is an excellent candidate for investigating the role of C-Type natriuretic Peptide in bone (1, 3).

A study comparing the measurement of CNP and NT-proCNP in human blood samples revealed that an NT-proCNP assay provides more consistent and reliable results, making it the preferred choice for clinical applications (5).

NT-proCNP can reliably be measured in serum and plasma samples with a conventional ELISA assay 

 BIOMEDICA – NT-proCNP ELISA (cat. no BI-20812) 

• Full validation  – following international quality guidelines
• For human and non-human samples (serum, plasma, cell culture, urine)
• Standard 96-well ELISA assay format- color coded and ready to use reagents
• Developed & manufactured by Biomedica  

 

NT-proCNP serves as a biomarker of linear growth  

Research studies in children has provided evidence showing a strong correlation between the levels of NT-proCNP in blood and the rate of linear growth throughout the entire growth process. In infancy, both males and females exhibit high levels of CNP and NT-proCNP, which subsequently decline in early childhood. However, during puberty, NT-proCNP levels rise again, followed by a decrease during adulthood. The peak levels of NT-proCNP align with the age at which peak height velocity occurs. As a result, the measurement of CNP syntheses through NT-proCNP in blood is highly indicative of linear growth in healthy children at all stages of development, making it an exceptional biomarker for monitoring linear growth (3, 6).

The role of NT-proCNP in bone diseases and genetic disorders

While the involvement of CNP in bone growth during development is well-established, its precise role in the adult skeleton remains uncertain. It is plausible that bone remodeling may contribute to increased plasma levels of CNP in children, given that CNP is expressed in osteoblasts and osteoclasts, which are the cells responsible for bone formation and bone degradation. Moreover, alternations in plasma CNP products have been linked to changes in bone turnover markers in adults (7),

CNP has emerged as a significant player in various types of skeletal dysplasia, a group of over 400 genetic disorders characterized by abnormal bone, joint, and cartilage development. Mutations in genes associated with CNP signaling have been identified in these conditions, including achondroplasia and hypochondroplasia, which are the most prevalent forms of dwarfism (8).

CNP has shown to be a promising therapeutic target for treating growth and skeletal disorders. Only recently, a variant of CNP was approved for treatment of achondroplasia (9). Ongoing clinical trials are currently investigating additional inhibitors of the CNP signaling pathway (10).

Related ELISA assay products

-NT-proANP (cat. no. BI-20492) – assay also suitable for preclinical applications (rat, mouse and other species) – 10µl sample volume

-NT-proBNP (cat. no. SK-1204) – CE-marked, for IVD use in EU

-NT-proBNP rat (cat. no. BI-1204R) – sample values provided

 

Literature

1. Circulating products of C-type natriuretic peptide and links with organ function in health and disease. Prickett TC, A Espiner E. Peptides. 2020. 132:170363.
2. C-type natriuretic peptide increases bone resorption in 1,25-dihydroxyvitamin D3-stimulated mouse bone marrow cultures. Holliday LS, Dean AD, Greenwald JE, Glucks SL. J Biol Chem. 1995. 11;270(32):18983-9.
3. Plasma C-Type Natriuretic Peptide: Emerging Applications in Disorders of Skeletal Growth. Espiner E, Prickett T, Olney R. Horm Res Paediatr. 2018;90(6):345-357.
4. Hormones and osteoporosis update. Effects of natriuretic peptides on endochondral bone growth. Clin Calcium. 2009. 19(7):1003-8. Yasoda A, Nakao K. Japanese. PMID: 19567998.
5. Comparative measurement of CNP and NT-proCNP in human blood samples: a methodological evaluation. Kuehnl A, Pelisek J, Bruckmeier M, Safi W, Eckstein HH. J Negat Results Biomed. 2013. 1;12:7.
6. Amino-terminal propeptide of C-type natriuretic peptide (NTproCNP) predicts height velocity in healthy children. Olney RC, Permuy JW, Prickett TC, Han JC, Espiner EA. Clin Endocrinol (Oxf). 2012. 77(3):416-22.
7. C-type natriuretic peptide forms in adult hyperthyroidism: correlation with thyroid hormones and markers of bone turnover. Schouten BJ, Prickett TC, Hunt PJ, Richards AM, Geffner ME, Olney RC, Espiner EA. Clin Endocrinol (Oxf). 2012. 76(6):790-6.
8. New developments in the management of achondroplasia. Högler W, Ward LM. Wien Med Wochenschr. 2020. 170(5-6):104-111.
9. Expanding horizons of achondroplasia treatment: current options and future developments. Fafilek B, Bosakova M, Krejci P. Osteoarthritis Cartilage. 2022. 30(4):535-544.
10. Emerging drug targets for achondroplasia. Savarirayan R. Expert Opin Ther Targets. 2022. 26(5):389-391.

 

 

 

 

We are happy to introduce our new brochure featuring our Quality Cytokine Assays “BIOMEDICA´s CYTOKINE ELISA kits”  

Product Highlights

• Full validation package
• Ready-to-use prediluted standards
• Color coded reagents & controls included
• Kit calibration with WHO international standard

QUALITY CYTOKINE ELISA KITS FOR AFFORDABLE & EFFICIENT RESEARCH

human Cytokine ELISA Kits  – developed & manufactured by Biomedica

• IL-6 (cat. no. BI-IL6) –  high sensitivity – detectable levels
• VEGF (cat. no. BI-VEGF) –  low sample volume – 10 µl – detecting all known VEGF isoforms
• ANGIOPOIETIN-2 (cat. no. BI-ANG2)    / optimized assay range 

 

Also available mouse/rat Angiopoietin-2 ELISA kit (cat. no. BI-ANG2MR) –  only 5µl sample volume.

 

 

ABOUT CYTOKINES

Cytokines are critical cell signaling molecules that play important roles in the immune response and many other physiological processes.  These proteins are produced by a wide variety of cell types that act on specific target cell receptors to trigger a range of biological responses.

Cytokines are involved in many aspects of the immune response, including inflammation, cell growth and differentiation, and cell death (apoptosis). Cytokines also regulate tissue repair and wound healing. Dysregulation of cytokine production and signaling may contribute to various diseases including autoimmune disorders, allergies, and cancer.

Some common cytokines include interleukins, tumor necrosis factor (TNF), and interferons.

Interleukin-6 (IL-6) is a pleiotropic cytokine that is largely involved in immunomodulatory processes. One of its roles is to respond to infections. As a pro-inflammatory cytokine, IL-6 activates the immune response, recruiting immune cells triggering B and T cell response. IL-6 dysregulation is associated with pathologies involving chronic inflammation, autoimmune diseases, and atherosclerosis. However, IL-6 also has beneficial anti-inflammatory and positive metabolic effects that are released during exercise coupled to a physical adaptation to intense training. In addition to its immune functions, IL-6 is involved in many other physiological processes, including bone remodeling and the nervous system.

Angiopoietin-2 (ANG2) is a protein that plays a critical role in regulating the development and remodeling of blood vessels, a process known as angiogenesis. ANG-2 is primarily produced by endothelial cells, which are cells that line the interior of blood vessels. Under normal physiological conditions the expression of ANG2 is low, which allows the blood vessels to undergo a rapid growth and remodeling. However, in pathological conditions, such as inflammation and cancer, ANG2 expression can be upregulated, leading to excessive angiogenesis and blood vessel destabilization.

Vascular endothelial growth factor (VEGF) is a protein mainly produced by endothelial cells. VEGF stimulates the growth and proliferation of endothelial cells, as well as the migration and differentiation of these cells into new blood vessels. VEGF plays a role in the development of cancer and certain ocular diseases, where it promotes the growth of new blood vessels that feed tumors or damage the retina.

There are several different isoforms of VEGF, each with different properties and functions. For example, VEGF-A is the most well-known cytokine of the VEGF family, and is primarily responsible for promoting angiogenesis. Other isoforms, such as VEGF-B and VEGF-C, have different roles in the body, such as regulating the growth of blood vessels in the heart and lymphatic system, respectively.

Related publications

Harnessing cytokines and chemokines for cancer therapy. Propper DJ, Balkwill FR. Nat Rev Clin Oncol. 2022 Apr;19(4):237-253. doi: 10.1038/s41571-021-00588-9. Epub 2022 Jan 7. PMID: 34997230.

Inflammatory Cytokines and Chemokines as Therapeutic Targets in Heart Failure. Hanna A, Frangogiannis NG. Cardiovasc Drugs Ther. 2020 Dec;34(6):849-863. doi: 10.1007/s10557-020-07071-0. Epub 2020 Sep 9. PMID: 32902739; PMCID: PMC7479403.

Around 7 out of 100,000 people are affected by primary malignant brain tumors. Glioblastoma multiforme (GBM) is the most common type of brain tumor. It grows rapidly and has a low prognosis of approximately 15 months. The  mean age after diagnosis is 63 years (1, 2). Despite significant progress in treatment, there remains a critical need for novel treatment options for GBM. 

Natural Malaria Substance – novel treatment option for brain tumors

Artemisinin, deriving from the plant Artemisia annua is a potent natural substance globally used to treat Malaria. It was discovered in the 1970s from Traditional Chinese Medicines and has since then saved millions of lives (3). Due to its cytotoxic properties it has not only been acknowledged for treating Malaria but also for its potential anticancer properties. With the discovery of Artemisinin and its derivates, researchers have been investigating the mechanisms on how these compounds work e.g. in inhibiting tumor cell proliferation, promoting apoptosis, disrupting caner invasion and metastasis, and preventing angiogenesis (4).

The natural malaria substance Artemisinin  –  a novel treatment option for brain tumors. 

In a recent study using genome-wide screening, researchers have identified mitochondrial function, particularly the biosynthesis of porphyrin, as a crucial pathway responsible for Artemisinin´s cytotoxicity. The potential of this novel treatment option for human brain tumors is currently under investigation (5, 6).

EZ4U / MTT Cell viability assay (cat. no. BI-5000) – BIOMEDICA

 

The Biomedica EZ4U cell viability and cytotoxicity assay was employed in mammalian cell lines in the described study:

A whole-genome scan for Artemisinin cytotoxicity reveals a novel therapy for human brain tumors.

The non-toxic and non-radioactive assay EZ4U / MTT Cell viability assay is based on the conversion of the yellow tetrazolium salt to coloured soluble formazan, which is produced by mitochondrial enzymes. The quantity of formazan produced is in direct proportion to the number of viable, living cells present.

Assay Characteristics

• Method: Reduction of tetrazolium salt to coloured formazan
• Sample type cell culture medium
• Sample size 200 μl / test, 10×96 tests
• Detection limit depending on cell lines Incubation time 2 – 5 h

 

Click here to download our EZ4U “easy for you” – brochure

Literature

1. Brain Tumors. Am J Med. McFaline-Figueroa JR, Lee EQ. 2018 Aug;131(8):874-882. doi: 10.1016/j.amjmed.2017.12.039. Epub 2018 Jan 31. PMID: 29371158.
2. Rising Incidence of Glioblastoma Multiforme in a Well-Defined Population. Grech N, Dalli T, Mizzi S, Meilak L, Calleja N, Zrinzo A Cureus. 2020 May 19;12(5):e8195. doi: 10.7759/cureus.8195. PMID: 32572354; PMCID: PMC7302718.
3. The discovery of artemisinin and the Nobel Prize in Physiology or Medicine. Su XZ, Miller LH. Sci China Life Sci. 2015 Nov;58(11):1175-9. doi: 10.1007/s11427-015-4948-7. PMID: 26481135; PMCID: PMC4966551.
4. Antitumor Research on Artemisinin and Its Bioactive Derivatives. Zhang Y, Xu G, Zhang S, Wang D, Saravana Prabha P, Zuo Z. Nat Prod Bioprospect. 2018 Aug;8(4):303-319. doi: 10.1007/s13659-018-0162-1. Epub 2018 Apr 9. PMID: 29633188; PMCID: PMC6102173.
5. A whole-genome scan for Artemisinin cytotoxicity reveals a novel therapy for human brain tumors. Taubenschmid-Stowers J, Orthofer M, Laemmerer A, Krauditsch C, Rózsová M, Studer C, Lötsch D, Gojo J, Gabler L, Dyczynski M, Efferth T, Hagelkruys A, Widhalm G, Peyrl A, Spiegl-Kreinecker S, Hoepfner D, Bian S, Berger W, Knoblich JA, Elling U, Horn M, Penninger JM. EMBO Mol Med. 2023 Mar 8;15(3):e16959. doi: 10.15252/emmm.202216959. Epub 2023 Feb 6. PMID: 36740985.
6. Achilles heel of glioblastoma. JLP Health and its partners discover combinatorial treatment option for incurable brain tumors. Press Release, JLP Health GmbH, Vienna, Austria, 7 February 2023.