Evaluating an in-house cell-based assay for detecting antibodies against muscle-specific tyrosine kinase in myasthenia gravis

Min Ju Kim, Seung Woo Kim, Mingi Kim, Young Chul Choi, Seung Min Kim, Ha Young Shin

Research output: Contribution to journalArticlepeer-review

Abstract

Background and Purpose Detecting antibodies against muscle-specific tyrosine kinase (MuSK Abs) is essential for diagnosing myasthenia gravis (MG). We applied an in-house cell-based assay (CBA) to detect MuSK Abs. Methods A stable cell line was generated using a lentiviral vector, which allowed the expression of MuSK tagged with green fluorescent protein in human embryonic kidney 293 (HEK293) cells. Serum and anti-human IgG antibody conjugated with red fluorescence were added. The presence of MuSK Abs was determined based on the fluorescence intensity and their colocal-ization in fluorescence microscopy. Totals of 218 serum samples collected from 177 patients with MG, 31 with other neuromuscular diseases, and 10 healthy controls were analyzed. The CBA results were compared with those of a radioimmunoprecipitation assay (RIPA) and an enzyme-linked immunosorbent assay (ELISA). Results The MuSK-HEK293 cell line stably expressed MuSK protein. The CBA detected MuSK Abs in 34 (19.2%) of 177 samples obtained from patients with MG and in none of the participants having other neuromuscular diseases or in the healthy controls. The clinical characteristics of the patients with MuSK MG determined based on the CBA were strongly corre-lated with known clinical features of MuSK MG. There was an almost perfect agreement between the results of the CBA and those of the RIPA (Cohen’s kappa=0.880, p<0.001) and ELISA (Cohen’s kappa=0.982, p<0.001). Conclusions The results of the in-house CBA showed excellent agreement with both the RIPA and ELISA. Our in-house CBA can be considered a reliable method for detecting MuSK Abs.

Original languageEnglish
Pages (from-to)400-408
Number of pages9
JournalJournal of Clinical Neurology (Korea)
Volume17
Issue number3
DOIs
Publication statusPublished - 2021 Jul

Bibliographical note

Funding Information:
This study was supported by a faculty research grant of Yonsei University College of Medicine (6-2016-0110).

Publisher Copyright:
© 2021 Korean Neurological Association.

All Science Journal Classification (ASJC) codes

  • Neurology
  • Clinical Neurology

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