Journal: Journal of Clinical Neurology (Seoul, Korea)

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

doi: 10.3988/jcn.2021.17.3.400

Figure Lengend Snippet: CBA to detect anti-MuSK Abs. When HEK293 cells expressing MuSK were incubated with (A) commercially available polyclonal anti-MuSK Abs or (B) sera from patients with MuSK-Ab-positive MG, anti-IgG antibody binding (red fluorescence) was observed along the cell surface, indicating the binding of MuSK Abs to MuSK. This colocalized with GFP-labeled MuSK expressed on the surface of HEK293 cells (green fluorescence). In contrast, (C) anti-human IgG antibody binding (red fluorescence) was not observed after adding the sera from the healthy control. Scale bar: 10 µm. CBA: cell-based assay, DAPI: 4′,6-diamidino-2-phenylindole, eGFP: enhanced green fluorescent protein, HEK293: human embryonic kidney 293, MG: myasthenia gravis, MuSK Abs: antibodies against muscle-specific tyrosine kinase.

Article Snippet: The cells were incubated with rabbit polyclonal anti-MuSK antibody (Invitrogen; 50 µg/mL final concentration) or sera from patients or controls (diluted 1:20) in Dulbecco's Modified Eagle's medium (DMEM) containing 1% bovine serum albumin and 20 mM HEPES, for 1 h at room temperature.

Techniques: Expressing, Incubation, Binding Assay, Fluorescence, Labeling, Cell Based Assay