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Analysis of Renal Function & Injury---β2-MG

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Human beta-2-microglobulin (β2-MG) is an 11.6 kDa non-glycosylated protein of 99 amino acids, which is the light chain of human leucocyte antigen-I (HLA-I) and can accumulate to cause serious dialysis-related amyloidosis (DRA) in long-term hemodialysis patients. It can be used for specific elimination of β2-MG from serum and can induce apoptosis of several types of tumor cells, and thus has great therapeutic potential. In addition to its physiological function, much clinical interest has been drawn to β2-MG as its increased serum levels and misfolding have been linked to a pathological condition.

 

Anti-Human β2-MG Monoclonal Antibodies 

A new generation of anti-β2-MG monoclonal antibodies, which was recently produced by CUSAg, makes possible the development of highly sensitive immunoassays. CUSAg antibodies are evaluated on different types of platforms: lateral-flow immunochromatogragphic assay (LFIA) and latex-enhanced turbidimetric immunoassay (LETIA).


Properties

Specification

Target species

Human

Host animal

Mice Balb/c

Cell line used for fusion

Sp2/0

Immunogen

Human β2-MG protein

Purification method, Purity

Protein G affinity chromatography

Presentation

MAb solution in PBS with 15 mM NaN3(pH 7.2)

Application

LFIA, LETIA

Catalog Number

CSB-DA003CmN①;

CSB-DA003CmN③;CSB-DA003CmN⑦

CSB-DA003CmN⑤;CSB-DA003CmN⑥

 

Calibration Curve

I. CLIA platform

β2-MG proteins specifically react with anti-β2-MG monoclonal antibodies(CSB-DA003CmN③and CSB-DA003CmN⑦)precoated onto latex beads to form insoluble complexs, resulting in turbidity increasing, and then the increasing of absorbance is detected by automatic biochemical analyzer. Our in-house assays have a linear detection range from 0-18 mg/L.The calibration curve was fitted according to the relationship between absorbance values and β2-MG concentrations (Fig.1).


 

Fig.1 Calibration curves for β2-MG in latex-enhanced turbidimetric immunoassay (LETIA) 

 

II. LETIA platform

A set of β2-MG calibrators with the concentration of 0、0.1、0.2、0.5、1、2 and 5 mg/L was detected on CUSAg LFIA platform using two anti-β2-MG monoclonal antibodies. The capture antibody was stripped on the nitrocellulose membrane, and the detection antibody was conjugated to colloidal gold. The best selected MAb combination for the development of semi-quantitative humanβ2-MG immunoassays is (capture-detection): CSB-DA003CmM⑥-CSB-DA003CmM⑤


Fig.2 Semi-quantitative detection of β2-MG protein in colloidal gold immunochromatogragphic assay


Clinical Comparison

A.LETIA platform
Anti-β2-MG monoclonal antibodies were also evaluated in medium-scale clinical trials with random blood samples from donates (n=44). Fig.3 showed that the correlation coefficient (r) is as high as 0.98 between in-house latex reagents and commercial β2-MG immunoassay. These results show good agreement between the two systems.

 

Fig.3  Clinical comparison of CUSAg β2-MG immunoassay and commercial diagnostic assay

 

 B.LFIA platform

20 samples from apparently healthy donors and patients were detected with the CUSAg LFIA β2-MG assay. As shown in Fig.4, the detection signals of T line were proportional to the concentration (0.06-31.3 mg/L) of the β2-MG in the sample tested by commercial kit.

 

Fig.4  Clinical comparison of CUSAg LETIA immunoassayand commercial immunoassays


β2-MG protein

A certain amount of excellent β2-MG protein (Cat:CSB-DP003A) is also offered by CUSAg. It can be used as calibrator in immunoassay.

 

Reference

1. P.E. EVRIN and L. WIBELL: The serum levels and urinary excretion of beta2-microglobulin in apparently healthy subjects. 1972; 29; 69-74

2. Kyung-sun Park, M.D., So-Young Kang, M.D., and Woo-In Lee, M.D.: Evaluation of two beta2-microglobulin reagents by turbidimetric immunoassay. 2007; 27; 414-9

3. Jae Ryung Shin, Seung Min Kim, Jung Sun Yoo, Ji Yoon Park, Seul Ki Kim, Joo Hee Cho, Kyung Hwan Jeong, Tae Won Lee and Chun Gyoo Ihm: Urinary excretion of beta2-microglobulin as a prognostic marker in immunoglobulin A in nephropathy. 2014; 29; 334-340

4. Pak Cheung R. Chan, Vathany Kulasingam and Bonny Lem-Ragosnig: Validating urinary measurement of beta-2-microglobulin with a Roche reagent kit designed for serum measurements. 2012; 45; 1533-1535

 

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