Related Experiment Video
Updated: Feb 13, 2026
Detection of Anti-MDA5 Autoantibodies Using HeLa Cells and Immunocytochemistry with Light Microscopy
Published on: October 31, 2025
Improved quantification of a commercial enzyme-linked immunosorbent assay kit for measuring anti-MDA5 antibody
Takahisa Gono1, Yuka Okazaki1, Akihiro Murakami2
1a Department of Allergy and Rheumatology , Nippon Medical School Graduate School of Medicine , Tokyo , Japan.
Objectives:
To compare the quantitative performance for measuring anti-MDA5 antibody titer of two enzyme-linked immunosorbent assay (ELISA) systems: an in-house ELISA and the commercial MESACUPTM anti-MDA5 test.
Methods:
Anti-MDA5 antibody titer was measured in sera from 70 patients with dermatomyositis using an in-house ELISA and the MESACUPTM anti-MDA5 test side-by-side. For the commercial ELISA kit, serum samples diluted 1:101 were used according to the manufacturer's protocol, but serial dilutions of sera were also examined to identify the optimal serum dilution for quantification.
Results:
The anti-MDA5 antibody titers measured by the in-house and commercial ELISAs were positively correlated with each other (r = 0.53, p = .0001), but the antibody titer measured by the commercial ELISA was less sensitive to change after medical treatment, and 37 (80%) of 46 anti-MDA5-positive sera had antibody titer exceeding the quantification range specified by the manufacturer (≥150 index). Experiments using diluted serum samples revealed that diluting the sera 1:5050 improved the quantitative performance of the MESACUPTM anti-MDA5 test, including a better correlation with the in-house ELISA results and an increased sensitivity to change.
Conclusion:
We improved the ability of the commercial ELISA kit to quantify anti-MDA5 antibody titer by altering its protocol.
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