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Evaluating an In-House Cell-Based Assay for Detecting Antibodies Against Muscle-Specific Tyrosine Kinase in Myasthenia Gravis

Authors
 Min Ju Kim  ;  Seung Woo Kim  ;  MinGi Kim  ;  Young Chul Choi  ;  Seung Min Kim  ;  Ha Young Shin 
Citation
 JOURNAL OF CLINICAL NEUROLOGY, Vol.17(3) : 400-408, 2021-07 
Journal Title
JOURNAL OF CLINICAL NEUROLOGY
ISSN
 1738-6586 
Issue Date
2021-07
Keywords
cell-based assay ; diagnosis ; muscle-specific tyrosine kinase ; myasthenia gravis
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 colocalization 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 correlated 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.
Files in This Item:
T202104703.pdf Download
DOI
10.3988/jcn.2021.17.3.400
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Neurology (신경과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Seung Min(김승민) ORCID logo https://orcid.org/0000-0002-4384-9640
Kim, Seung Woo(김승우) ORCID logo https://orcid.org/0000-0002-5621-0811
Shin, Ha Young(신하영) ORCID logo https://orcid.org/0000-0002-4408-8265
Choi, Young Chul(최영철) ORCID logo https://orcid.org/0000-0001-5525-6861
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/186823
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