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  • Qualitative and Quantitative Assay for Detection of Circulating Autoantibodies against Human Aortic Antigen.

Qualitative and Quantitative Assay for Detection of Circulating Autoantibodies against Human Aortic Antigen.

Bio-protocol (2017-07-05)
Brent Veerman, Ritu Chakravarti
ABSTRACT

Increased amount of autoantibodies in human sera are the hallmark of autoimmune diseases ( Wang et al., 2015 ). In case of known antigen, detection of autoantibodies is done using laboratory based methods. However, in most autoimmune diseases, knowledge of self-antigen is still vague. We have developed an ELISA-based quantitative assay to detect the presence of autoantibodies as well as to measure the circulating autoantibodies in the sera of patients suffering from large vessel vasculitis (LVV), an autoimmune disease ( Chakravarti et al., 2015 ). Using this assay, we detected the increase in anti-aortic antibodies in LVV patient's sera. We have further verified the results by independent biochemical techniques and found the specificity to be > 94% ( Chakravarti et al., 2015 ). This method can be uniquely modified to suit any autoimmune, in particular organ specific, disease and thus has wider applications in the detection and quantification of autoantibodies.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium orthovanadate, ≥90% (titration)
Sigma-Aldrich
Sodium chloride, ACS reagent, ≥99.0%
Sigma-Aldrich
β-Glycerophosphate disodium salt hydrate, BioUltra, suitable for cell culture, suitable for plant cell culture, ≥99% (titration)