Download biomedica product listHuman Vanin-1 (urine) ELISA | BI-VAN1U
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Method
Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
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Sample type
Urine
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Sample volume
10 µl / well
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Assay time
4 h / 30 min
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Sensitivity
9.6 pmol/l (= 500 pg/ml)
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Standard range
0 – 1 200 pmol/l (= 0 – 62 500 pg/ml)
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Conversion factor
1 pmol/l = 52.07 pg/ml (MW: 52.07 kDa)
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Precision
In-between-run (n=9): ≤ 7 % CV
Within-run (n=3): ≤ 5 % CV
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Specificity
Endogenous and recombinant human Vanin-1.
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Cross-reactivity
No cross-reactivity with mouse Vanin-1.
Potentially cross-reactive with Vanin-1 from various monkey species.
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Use
Research use only
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Validation Data
See validation data tab for: precision, accuracy, dilution linearity, values for healthy donors, etc
Human Vanin-1 ELISA Product Overview
The human Vanin-1 ELISA kit is a 4.5 h, 96-well sandwich ELISA for the quantitative determination of Vanin-1 in urine.
The Vanin-1 ELISA kit uses highly purified, epitope mapped antibodies.
Human Vanin-1 ELISA Assay Principle
The Vanin ELISA kit is a sandwich enzyme immunoassay for the quantitative determination of Vanin-1 in human urine.
The figure below explains the principle of the Vanin-1 sandwich ELISA:
Capture antibody: polyclonal sheep anti-human Vanin-1 antibody
Detection antibody: polyclonal sheep anti-human Vanin-1 antibody, HRP-labeled
Target antigen: human Vanin-1
In a first step, assay buffer is pipetted into the wells of the microtiter strips. Thereafter, standard/control/sample and detection antibody (polyclonal sheep anti-human Vanin-1-HRP) are pipetted into the wells, which are pre-coated with polyclonal sheep anti-human Vanin-1 antibody. Vanin-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. In the washing step, all non-specific unbound material is removed. In a next step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of Vanin-1 present in the sample. This color change is detectable with a standard microplate reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated using the values obtained from the standards. The concentration of Vanin-1 in the sample is determined directly from the dose response curve.
Human Vanin-1 ELISA Typical Standard Curve
The figure below shows a typical standard curve for the Vanin-1 ELISA assay. The immunoassay is calibrated against recombinant full-length human Vanin-1 protein:
Human Vanin-1 ELISA Kit Components
|
Contents |
Description |
Quantity |
|
PLATE |
Polyclonal sheep anti-human Vanin-1 antibody pre-coated microtiter strips in a strip holder, packed in an aluminum bag with desiccant |
12 x 8 tests |
|
WASHBUF |
Wash buffer concentrate 20x, natural cap |
1 x 50 ml |
|
STD |
Standards 1-7, (0; 37.5; 75; 150; 300; 600; 1200 pmol/l), recombinant human Vanin-1 in buffer, white caps, lyophilized |
7 vials |
|
CTRL |
Control A and B, yellow cap, lyophilized, exact concentrations see labels |
2 vials |
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ASYBUF |
Assay buffer, red cap, ready to use |
1 x 7 ml |
|
CONJ |
Conjugate (polyclonal sheep anti-human Vanin-1-HRP), amber bottle, amber cap, ready to use |
1 x 6 ml |
|
SUB |
Substrate (TMB solution), amber bottle, blue cap, ready to use |
1 x 13 ml |
|
STOP |
Stop solution, white cap, ready to use |
1 x 7 ml |
Storage instructions: all reagents of the Vanin-1 (urine) ELISA kit are stable at 4°C (2-8°C) until the expiry date stated on the label of each reagent.
Urine is suitable for use in this Vanin-1 ELISA. Do not change sample type during studies. We recommend duplicate measurements for all samples, standards and controls. The sample collection and storage conditions listed are intended as general guidelines.
Urine
Aseptically collect the first urine of the day (mid-stream), voided directly into a sterile container. Centrifuge to remove particulate matter, assay immediately or aliquot and store at -25°C or lower. Samples can undergo at least four freeze-thaw cycles.
Reagent Preparation
Wash Buffer
|
1. |
Bring the WASHBUF concentrate to room temperature. Crystals in the buffer concentrate will dissolve at room temperature. |
|
2. |
Dilute the WASHBUF concentrate 1:20, e.g. 50 ml WASHBUF + 950 ml distilled or deionized water. Only use diluted WASHBUF when performing the assay. |
The diluted WASHBUF is stable up to one month at 4°C (2-8°C).
Standards
|
1. |
Pipette 200 µl of distilled or deionized water into each standard (STD) and control (CTRL) vial. The exact concentration is printed on the label of each vial. |
|
2. |
Leave at room temperature (18-26°C) for 10 min. Vortex gently. |
Reconstituted STDs and the CTRL are stable for three hours at room temperature (18-26°C). STDs and CTRLs are stable -25°C or lower until expiry date stated on the label and can be subjected to up to three freeze-thaw cycles.
Sample Preparation
Bring samples to room temperature and mix samples gently to ensure the samples are homogeneous. We recommend duplicate measurements for all samples.
Samples for which the OD value exceeds the highest point of the standard range can be diluted with assay buffer.
Human Vanin-1 ELISA Assay Protocol
Read the entire protocol before beginning the assay.
|
1. |
Bring samples and reagents to room temperature (18-26°C). |
|
2. |
Mark positions for STD/CTRL/SAMPLE (standard/control/sample) on the protocol sheet. |
|
3. |
Take microtiter strips out of the aluminum bag. Store unused strips with desiccant at 4°C in the aluminum bag. Strips are stable until expiry date stated on the label. |
|
4. |
Pipette 50 µl ASYBUF (assay buffer, red cap) into each well. |
|
5. |
Add 10 µl STD/CTRL/SAMPLE into the respective wells. |
|
6. |
Add 50 µl CONJ (conjuagate, amber cap) into each well. Swirl gently. |
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7. |
Cover tightly and incubate for 4 hours at room temperature (18-26°C) in the dark. |
|
8. |
Aspirate and wash wells 5 x with 300 µl diluted WASHBUF (wash buffer). After the final wash, remove the remaining WASHBUF by strongly tapping plate against a paper towel. |
|
12. |
Add 100 µl SUB (substrate, blue cap) into each well. |
|
13. |
Incubate for 30 min at room temperature (18-26°C) in the dark. |
|
14. |
Add 50 µl STOP (stop solution, white cap) into each well. |
|
15. |
Measure absorbance immediately at 450 nm with reference 630 nm, if available. |
Calculation of Results
Read the optical density (OD) of all wells on a plate reader using 450 nm wavelength (reference wavelength 630 nm). Construct a standard curve from the absorbance read-outs of the standards using commercially available software capable of generating a four-parameter logistic (4-PL) fit. Alternatively, plot the standards’ concentration on the x-axis against the mean absorbance for each standard on the y-axis and draw a best fit curve through the points on the graph. Curve fitting algorithms other than 4-PL have not been validated and will need to be evaluated by the user.
Obtain sample concentrations from the standard curve. If required, pmol/l can be converted into pg/ml by applying a conversion factor (1 pg/ml = 0.0192 pmol/l (MW: 52.07 kDa)). Respective dilution factors must be considered when calculating the final concentration of the sample.
The quality control protocol supplied with the kit shows the results of the final release QC for each kit at production date. Data for OD obtained by customers may differ due to various influences including the normal decrease of signal intensity throughout shelf life. However, this does not affect validity of results as long as an OD of 1.5 or higher is obtained for the standard with the highest concentration and the control value is in range (target range see label).
Depending on the study population and the outcome measures applied, urinary Vanin-1 values should be normalized to urinary Creatinine values (Tang et al., 2015).
Normalisation of urinary biomarkers to creatinine for clinical practice and research – when and why.
Tang, K.W.A., Toh, Q.C., Teo, B.W., 2015. Singapore Med J 56, 7–10.
PMID: 25640093; PMCID: PMC4325562
Vanin-1 Protein
Vanin-1 is a GPI-anchored glycoprotein of 513 amino acids consisting of a base domain and an enzymatic nitrilase domain (Boersma et al., 2014). The ectoenzyme catalyzes the hydrolysis of pantetheine to pantothenic acid (vitamin B5) and cyteamine and thus, is involved in the regulation of oxidative stress and inflammation (Maras et al., 1999). Vanin-1 has a broad tissue expression with the highest levels being observed in kidney tubular epithelial cells (Pitari et al., 2000).The GPI anchor of Vanin-1 can be cleaved by a yet unknown mechanism, resulting in Vanin-1 being shed into the extracellular space.
|
Molecular weight |
52.07 kDa |
|
Cellular localization |
Extracellular, plasma membrane |
|
Post-translational modifications |
Glycosylation, lipidation (GPI-anchor) |
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Sequence similarities |
Member of the Vanin family of proteins, sequence similarities with biotinidase family |
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Alternative names |
Vascular non-inflammatory molecule 1, Tiff66, pantetheine hydrolase, pantetheinase, VNN1, HDLCQ8 |
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Entrez/NCBI ID |
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Genecards |
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OMIM |
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Protein Atlas |
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Uniprot ID |
Vanin-1 Function
Vanin-1 is an epithelial ectoenzyme activating the conversion of pantetheine into pantothenic acid (vitamin B5) and cysteamine (Pitari et al., 2000). It has been suggested that the release of cysteamine by Vanin-1 promotes oxidative tissue damage and inflammation by inhibiting the activity of antioxidants like superoxide dismutase (SOD) and glutathione (GSH) (Hosohata et al., 2011; Saghaei et al., 2012). Indeed, Vanin-1 knockout mice have elevated stores of GSH and are more resistant to oxidative injury induced by whole-body gamma irradiation (Berruyer et al., 2004). On the other hand, several reports indicate that Vanin-1 might also act as tissue sensor for oxidative stress. In mice, antioxidant response-like elements could be identified in the promotor region of Vanin-1, which enhance the expression of Vanin-1 in the presence of oxidative stress (Berruyer et al., 2004). Similarly, Vanin-1 expression was shown to be upregulated in a human proximal tubular cell line after exposure to organic solvents (Hosohata et al., 2011). After renal ischemia-reperfusion in rats, a model involving oxidative tissue damage, renal Vanin-1 expression was also found to be upregulated (Yoshida et al., 2002).
The highest levels of Vanin-1 expression could be assigned to renal tubular epithelial cells, while no expression is detectable in glomeruli (Hosohata et al., 2011; Pitari et al., 2000). Hence, Vanin-1 released from renal cells could be detectable in urine. In a study aimed to identify biomarkers for renal tubular injury, Hosohata and colleagues could indeed show in a rat model of nephrotoxicant-induced injury that Vanin-1 is upregulated in renal tubules earlier than other markers and shed into urine (Hosohata et al., 2011). Subsequent studies further verified the validity of Vanin-1 as an early biomarker of renal tubular damage in drug-induced acute kidney injury (Hosohata et al., 2012, 2016a), obstructive nephropathy (Washino et al., 2019) and hydronephrosis (Hosohata et al., 2018), diabetic nephropathy (Fugmann et al., 2011), renal injury in experimental colitis (Hosohata et al., 2014) and spontaneously hypertensive rats under high salt intake (Hosohata et al., 2016b; Washino et al., 2018). Of note, Vanin-1 seems to have superior predictive value for acute kidney injury than established markers KIM-1, NGAL, or NAG (Fugmann et al., 2011; Hosohata, 2016; Hosohata et al., 2011).
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Nephrology
- Acute kidney injury (Hosohata et al., 2016a)
- Diabetic nephropathy (Fugmann et al., 2011)
- Drug-induced acute kidney injury (Hosohata et al., 2016a)
- Hydronephrosis (Hosohata et al., 2018), obstructive nephropathy (Washino et al., 2019)
Literature
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Vanin-1-/- mice exhibit a glutathione-mediated tissue resistance to oxidative stress.
Berruyer, C., Martin, F.M., Castellano, R., Macone, A., Malergue, F., Garrido-Urbani, S., Millet, V., Imbert, J., Duprè, S., Pitari, G., Naquet, P., Galland, F., 2004. Mol. Cell. Biol. 24, 7214–7224.
PMID:15282320; PMCID: PMC479710 -
The structure of vanin 1: a key enzyme linking metabolic disease and inflammation.
Boersma, Y.L., Newman, J., Adams, T.E., Cowieson, N., Krippner, G., Bozaoglu, K., Peat, T.S., 2014. Acta Crystallogr. D Biol. Crystallogr. 70, 3320–3329.
PMID:25478849 -
Fugmann, T., Borgia, B., Révész, C., Godó, M., Forsblom, C., Hamar, P., Holthöfer, H., Neri, D., Roesli, C., 2011. Kidney Int. 80, 272–281.
PMID:21544065 -
Vanin-1 in Renal Pelvic Urine Reflects Kidney Injury in a Rat Model of Hydronephrosis.
Hosohata, K., Jin, D., Takai, S., Iwanaga, K., 2018. Int J Mol Sci 19.
PMID:30332759; PMCID: PMC6214032 -
Role of Oxidative Stress in Drug-Induced Kidney Injury.
Hosohata, K., 2016. Int J Mol Sci 17.
PMID:27809280; PMCID: PMC5133827 -
Hosohata, K., Washino, S., Kubo, T., Natsui, S., Fujisaki, A., Kurokawa, S., Ando, H., Fujimura, A., Morita, T., 2016a. Toxicology 359–360, 71–75.
PMID:27317936 -
Hosohata, K., Yoshioka, D., Tanaka, A., Ando, H., Fujimura, A., 2016b. Hypertens. Res. 39, 19–26.
PMID:26376947 -
Early detection of renal injury using urinary vanin-1 in rats with experimental colitis.
Hosohata, K., Ando, H., Fujimura, A., 2014. J Appl Toxicol 34, 184–190.
PMID:23307618 -
Urinary Vanin-1 As a Novel Biomarker for Early Detection of Drug-Induced Acute Kidney Injury.
Hosohata, K., Ando, H., Fujimura, A., 2012. Journal of Pharmacology and Experimental Therapeutics 341, 656–662.
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Vanin-1: a potential biomarker for nephrotoxicant-induced renal injury.
Hosohata, K., Ando, H., Fujiwara, Y., Fujimura, A., 2011. Toxicology 290, 82–88.
PMID:21907259 -
Is pantetheinase the actual identity of mouse and human vanin-1 proteins?
Maras, B., Barra, D., Duprè, S., Pitari, G., 1999. FEBS Letters 461, 149–152.
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Pitari, G., Malergue, F., Martin, F., Philippe, J.M., Massucci, M.T., Chabret, C., Maras, B., Duprè, S., Naquet, P., Galland, F., 2000. FEBS Letters 483, 149–154.
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Effects of captopril on the cysteamine-induced duodenal ulcer in the rat.
Saghaei, F., Karimi, I., Jouyban, A., Samini, M., 2012. Exp. Toxicol. Pathol. 64, 373–377.
PMID:21036019 -
Washino, S., Hosohata, K., Oshima, M., Okochi, T., Konishi, T., Nakamura, Y., Saito, K., Miyagawa, T., 2019. Int J Mol Sci 20.
PMID:30791405; PMCID: PMC6412925 -
Washino, S., Hosohata, K., Jin, D., Takai, S., Miyagawa, T., 2018. Clin. Exp. Pharmacol. Physiol. 45, 261–268.
PMID:29027259 -
Monitoring changes in gene expression in renal ischemia-reperfusion in the rat.
Yoshida, T., Kurella, M., Beato, F., Min, H., Ingelfinger, J.R., Stears, R.L., Swinford, R.D., Gullans, S.R., Tang, S.-S., 2002. Kidney Int. 61, 1646–1654.
PMID:11967014
PMID:15282320; PMCID: PMC479710All Biomedica ELISAs are validated according to international FDA/ICH/EMEA guidelines. For more information about our validation guidelines, please refer to our quality page and published validation guidelines and literature.
- ICH Q2(R1) Validation of Analytical Procedures: Text and Methodology.
- EMEA/CHMP/EWP/192217/2009 Guideline on bioanalytical method validation.
- Bioanalytical Method Validation, Guidance for Industry, FDA, May 2018
Calibration
The Vanin-1 (urine) immunoassay is calibrated against recombinant full-length human Vanin-1 protein (O95497 (Uniprot ID)).
Human Vanin-1 ELISA Detection Limit & Sensitivity
To determine the sensitivity of the Vanin-1 (urine) ELISA, experiments measuring the lower limit of detection (LOD) and the lower limit of quantification (LLOQ) were conducted.
The LOD, also called the detection limit, is the lowest point at which a signal can be distinguished above the background signal, i.e. the signal that is measured in the absence of Vanin-1, with a confidence level of 99%. It is defined as the mean back calculated concentration of standard 1 (0 pmol/l of Vanin-1, five independent measurements) plus three times the standard deviation of the measurements.
The LLOQ, or sensitivity of an assay, is the lowest concentration at which an analyte can be accurately quantified. The criteria for accurate quantification at the LLOQ are an analyte recovery between 75 and 125% and a coefficient of variation (CV) of less than 25%. To determine the LLOQ, standard 2, i.e. the lowest standard containing Vanin-1, is diluted, measured five times and its concentration is back calculated. The lowest dilution, which meets both criteria, is reported as the LLOQ.
The following values were determined for the Vanin-1 ELISA:
|
LOD |
9.6 pmol/l |
|
LLOQ |
38 pmol/l |
Human Vanin-1 ELISA Precision
The precision of an ELISA is defined as its ability to measure the same concentration consistently within the same experiments carried out by one operator (within-run precision or repeatability) and across several experiments using the same samples but conducted by several operators using different ELISA lots (in-between-run precision or reproducibility).
Within-Run Precision
Within-run (intra-assay) precision was assessed by measuring two samples of known concentrations three times within one Vanin-1 human urinary ELISA kit lot by one operator.
|
ID |
n |
Mean Vanin-1 [pmol/l] |
SD [pmol/l] |
CV (%) |
|
Sample 1 |
3 |
74 |
3 |
5 |
|
Sample 2 |
3 |
606 |
24 |
4 |
In-Between-Run Precision
In progress.
Human Vanin-1 ELISA Accuracy
The accuracy of an ELISA is defined as the precision with which it can recover samples of known concentrations.
The recovery of the Vanin-1 human urinary ELISA was measured by adding recombinant Vanin-1 to human urine samples containing a known concentration endogenous Vanin-1. The % recovery of the spiked concentration was calculated as the percentage of measured compared over the expected value.
The table shows the summary of the recovery experiments in the human Vanin-1 (urine) ELISA in human urine samples:
|
% Recovery |
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|
+120 pmol/l |
+600 pmol/l |
||||
|
Sample matrix |
n |
Mean |
Range |
Mean |
Range |
|
Urine |
6 |
81 |
73-92 |
93 |
86-99 |
Data showing % recovery of recombinant Vanin-1 in human urine samples:
|
Vanin-1 [pmol/l] |
% Recovery |
|||||
|
Sample matrix |
ID |
Reference |
+ 120 pmol/l |
+ 600 pmol/l |
+ 120 pmol/l |
+ 600 pmol/l |
|
Urine |
u1 |
44 | 149 | 588 | 92 | 94 |
|
Urine |
u2 |
62 | 156 | 597 | 84 | 94 |
|
Urine |
u3 |
104 | 157 | 595 | 79 | 90 |
|
Urine |
u4 |
19 | 99 | 527 | 73 | 86 |
|
Urine |
u5 |
229 | 232 | 673 | 79 | 93 |
|
Urine |
u6 |
86 | 147 | 637 | 79 | 99 |
|
Mean |
81 | 93 | ||||
|
Min |
73 | 86 | ||||
|
Max |
92 | 99 | ||||
Human Vanin-1 ELISA Dilution Linearity & Parallelism
Tests of dilution linearity and parallelism ensure that both endogenous and recombinant samples containing Sclerostin behave in a dose dependent manner and are not affected by matrix effects. Dilution linearity assesses the accuracy of measurements in diluted clinical samples spiked with known concentrations of recombinant analyte. By contrast, parallelism refers to dilution linearity in clinical samples and provides evidence that the endogenous analyte behaves in same way as the recombinant one. Dilution linearity and parallelism are assessed for each sample type and are considered acceptable if the results are within ± 20% of the expected concentration.
Dilution Linearity
Dilution linearity was assessed by serially diluting human urine samples spiked with 600 pmol/l recombinant Vanin-1 with assay buffer.
The table below shows the mean recovery and range of serially diluted recombinant Vanin-1 in urine:
|
% Recovery of recombinant Vanin-1 in diluted samples |
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|
|
1+1 |
1+3 |
1+7 |
||||
|
Sample matrix |
n |
Mean |
Range |
Mean |
Range |
Mean |
Range |
|
Urine |
6 |
94 |
91-97 |
91 | 88-93 | 80 |
78-82 |
Data showing dilution linearity of human urine samples containing recombinant Vanin-1:
|
Vanin-1 [pmol/l] |
Recovery (%) |
|||||||
|
Sample matrix |
ID |
Reference |
1+1 |
1+3 |
1+7 |
1+1 |
1+3 |
1+7 |
|
Urine |
u1 | 629 | 294 | 126 | 60 | 94 | 80 | 76 |
|
Urine |
u2 | 564 | 297 | 141 | 62 | 105 | 100 | 87 |
|
Urine |
u3 | 588 | 279 | 130 | 59 | 95 | 88 | 81 |
|
Urine |
u4 | 609 | 286 | 140 | 63 | 94 | 92 | 82 |
|
Urine |
u5 | 597 | 290 | 139 | 59 | 97 | 93 | 79 |
|
Urine |
u6 | 642 | 293 | 146 | 62 | 91 | 91 | 78 |
|
|
|
|
|
|
Mean |
94 | 91 | 80 |
|
|
|
|
|
|
Min |
91 | 88 | 78 |
|
|
|
|
|
|
Max |
97 | 93 | 82 |
Parallelism
Parallelism was assessed by serially diluting urine samples containing endogenous Vanin-1 with assay buffer.
The table below shows the mean recovery and range of serially diluted endogenous Vanin-1 in human urine:
| % Recovery of endogenous Vanin-1 in diluted samples | |||||||
|
1+1 |
1+3 |
1+7 |
|||||
|
Sample matrix |
n |
Mean |
Range |
Mean |
Range |
Mean |
Range |
|
Urine |
6 |
94 |
85-100 |
92 |
79-98 |
86 |
69-99 |
Data showing dilution linearity of human urine samples containing endogenous Vanin-1:
|
Vanin-1 [pmol/l] |
% Recovery |
|||||||
|
Sample matrix |
ID |
Reference |
1+1 |
1+3 |
1+7 |
1+1 |
1+3 |
1+7 |
|
Urine |
u1 |
1188 | 593 | 291 | 146 | 100 | 98 | 99 |
|
Urine |
u2 |
717 | 357 | 169 | 83 | 100 | 94 | 93 |
|
Urine |
u3 |
1009 | 431 | 219 | 102 | 85 | 87 | 81 |
|
Urine |
u4 |
623 | 267 | 123 | 54 | 86 | 79 | 69 |
| Urine | u5 |
657 |
313 | 161 |
75 |
95 |
98 |
91 |
| Urine | u6 |
607 |
289 | 143 |
65 |
95 |
94 |
85 |
|
|
Mean |
94 | 92 | 86 | ||||
|
|
Min |
85 | 79 | 69 | ||||
|
|
Max |
100 | 98 | 99 | ||||
Human Vanin-1 ELISA Specificity
The specificity of an ELISA is defined as its ability to exclusively recognize the analyte of interest.
The specificity of the Vanin-1 (urine) ELISA was shown by characterizing both the capture and the detection antibodies through epitope mapping. In addition, the specificity of the ELISA was established through competition experiments, which measure the ability of the antibodies to exclusively bind Vanin-1.
This assay recognizes recombinant and endogenous human Vanin-1.
Epitope Mapping
Antibody binding sites were determined by epitope mapping using microarray analysis (Pepperprint GmbH). The capture antibody binds to a single linear epitope located in the nitrilase domain of Vanin-1. The detection antibody binds to several epitopes distributed over the entire Vanin-1 molecule. For more information please contact info@bmgrp.com.
Competition of Signal
Competition experiments were carried out by pre-incubating human samples containing endogenous Vanin-1 with an excess of capture antibody (AB). The concentration measured in this mixture was then compared to a reference value, which was obtained from the same sample without the pre-incubation step. Mean competition in urine samples was 100%.
|
Vanin-1 [pmol/l] |
|
|||
|
Sample matrix |
ID |
Reference |
Reference + capture AB |
% Competition |
|
Urine |
u1 |
1253 | 29 |
98 |
|
Urine |
u2 |
980 | 0 |
100 |
|
Urine |
u3 |
107 | 0 |
100 |
|
Urine |
u4 |
385 | 0 |
100 |
|
Urine |
u5 |
869 | 0 |
100 |
|
Urine |
u6 |
238 | 0 |
100 |
|
Mean |
100 |
|||
Isoforms
There are no isoforms of Vanin-1 known.
Cross Reactivity
Potentially cross-reactive with Vanin-1 from various monkey species. No cross-reactivity with mouse Vanin-1.
Sample Stability
Freeze-thaw Stability
The freeze-thaw stability of endogenous Vanin-1 was tested by comparing Vanin-1 measurements in urine samples that had undergone four freeze-thaw cycles.
For freeze-thaw experiments, a set of human urine samples was aliquoted and freeze-thaw stressed. Samples can undergo at least four freeze-thaw cycles. The mean recovery of sample concentrations stressed by four freeze-thaw cycles is 96%.
Vanin-1 concentrations of samples after freeze-thaw (F/T) cycles:
|
Vanin-1 [pmol/l] |
Recovery (%) 4x F/T vs Reference |
||||||
|
Sample matrix |
ID |
Reference |
1x |
2x |
3x |
4x |
|
|
Urine |
u1 |
108 | 98 | 100 | 86 | 86 | 80 |
|
Urine |
u2 |
29 |
26 |
26 |
29 |
29 |
100 |
|
Urine |
u3 |
1218 |
1172 |
1186 |
1051 |
1210 |
99 |
|
Urine |
u4 |
872 |
906 |
815 |
786 |
826 |
95 |
|
Urine |
u5 |
569 |
670 |
552 |
603 |
593 |
104 |
|
Urine |
u6 |
128 |
170 |
176 |
128 |
100 |
100 |
|
Mean |
96 |
||||||
Benchtop Stability
The benchtop stability of endogenous Vanin-1 was tested by comparing Vanin-1 measurements in urine samples that had been stored at different temperatures.
For the assessment of the benchtop stability, a set of human urine samples was aliquoted and stored at -25°C, at room temperature or at 4°C. Samples can be stored for at least three hours at room temperature as well as overnight at 4°C . The mean recovery of sample concentrations after three hours at room temperature is 99%. The mean recovery of sample concentrations after overnight storage at 4°C is 87%.
Vanin-1 concentrations of samples stored at -25°C (reference), at room temperature (RT) or overnight (ON) at 4°C:
|
Vanin-1 [pmol/l] |
Recovery (%) vs reference |
|||||||
|
Sample matrix |
ID |
Reference |
1 h RT |
3 h RT |
ON 4°C |
1 h RT |
3 h RT |
ON 4°C |
|
Urine |
u1 |
99 | 92 | 97 | 84 | 92 | 97 | 85 |
|
Urine |
u2 |
25 |
26 | 27 | 27 | 102 | 107 | 107 |
|
Urine |
u3 |
1158 |
1259 | 1191 | 1297 | 109 | 103 | 112 |
|
Urine |
u4 |
963 |
1000 | 806 | 899 | 104 | 84 | 93 |
|
Urine |
u5 |
752 |
707 | 600 | 536 | 94 | 80 | 71 |
|
Urine |
u6 |
126 |
100 | 156 | 67 | 80 | 124 | 53 |
|
Mean |
97 |
99 |
87 |
|||||
Sample Values
Vanin-1 Values in Apparently Healthy Individuals
To provide values for Vanin-1 in apparently healthy individuals, a panel of urine samples was tested.
A summary of the results is shown below:
| Vanin-1 [pmol/l] | |||||
|
Sample matrix |
n |
Mean |
Range |
SD |
Median |
|
Urine |
27 |
196 |
3-963 |
197 |
116 |
Normalized Vanin-1 Values in Apparently Healthy Individuals
Vanin-1 values in urine samples from apparently healthy individuals were converted from pmol/l into pg/ml (conversion factor: 1 pg/ml= 0.0192 pmol/l) and normalized to Creatinine values.
A summary of the normlized results is shown below:
| Vanin-1 [pg/mg Creatinine] | |||||
|
Sample matrix |
n |
Mean |
Range |
SD |
Median |
|
Urine |
27 |
1244 | 77-2813 | 751 | 1131 |
It is recommended to establish the normal range for each laboratory.
Vanin-1 Values in Individuals with Kidney Disease
In addition to samples from apparently healthy donors, a panel of samples from a kidney disease cohort was tested.
A summary of the results is shown below:
| Vanin-1 [pmol/l] | ||||||
|
Cohort |
Sample matrix |
n |
Mean |
Range |
SD |
Median |
|
Apparenty helathy |
Urine |
27 |
196 |
3-963 |
197 |
116 |
|
Kidney disease |
Urine |
24 |
589 |
57-2375 |
561 |
360 |
Normalized Vanin-1 Values in Individuals with Kidney Disease
Vanin-1 values in urine samples from individuals with kidney disease were converted from pmol/l into pg/ml (conversion factor: 1 pg/ml= 0.0192 pmol/l) and normalized to Creatinine values.
A summary of the normlized results is shown below:
| Vanin-1 [pg/mg Creatinine] | ||||||
|
Cohort |
Sample matrix |
n |
Mean |
Range |
SD |
Median |
|
Apparenty healthy |
Urine |
27 |
1244 |
77-2813 |
751 |
1131 |
|
Kidney disease |
Urine |
24 |
5690 |
1101-28764 |
6239 |
3280 |
- The Usefulness of Vanin-1 and Periostin as Markers of an Active Autoimmune Process or Renal Fibrosis in Children with IgA Nephropathy and IgA Vasculitis with Nephritis-A Pilot Study. Mizerska-Wasiak M, Płatos E, Cichoń-Kawa K, Demkow U, Pańczyk-Tomaszewska M. J Clin Med. 2022 Feb 25;11(5):1265. doi: 10.3390/jcm11051265. PMID: 35268356; PMCID: PMC8911128.
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Urinary vanin-1 for predicting acute pyelonephritis in young children with urinary tract infection: a pilot study. Krzemień G, Pańczyk-Tomaszewska M, Górska E, Szmigielska A. Biomarkers. 2021. 26(4):318-324.
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Development of an immunoassay that reveals altered uninary Vanin-1 in human with kidney disease.Wallwitz, J., Eichinger, B., Berg, G., Gadermaier, E., Himmler, G. 2018. Nephrology Dialysis Transplantation, Volume 33, Issue suppl_1, Page i126. ERA-EDTA Poster 2018 Vanin
