Total MuSK autoantibody levels were assessed by RIA (RSR, UK) according to the manufacturer’s instructions. MuSK IgG4 levels were measured by ELISA. To this end, ELISA plates (Microlon, catalog number 655092, Greiner, Austria) were coated with 1 µg/mL MuSK extracellular domain (ECD) produced in mammalian cells (a kind gift of Dr. Bernard Rees-Smith, RSR, UK). Bound MuSK antibodies were detected with mouse anti-human IgG4:HRP (1:3,500, catalog number MCA2098P, AbD Serotec, Germany). Samples were incubated with substrate containing 3,3′,5,5′-tetramethylbenzidine, and absorbance at 450 nm was measured using a VictorX3 plate reader (PerkinElmer, USA).
Koneczny I., Mané-Damas M., Zong S., De Haas S., Huda S., van Kruining D., Damoiseaux J., De Rosa A., Maestri M., Guida M., Molenaar P., Van Damme P., Fichtenbaum A., Perkmann T., De Baets M., Lazaridis K., Zouvelou V., Tzartos S., Ricciardi R., Losen M, & Martinez-Martinez P. (2024). A retrospective multicenter study on clinical and serological parameters in patients with MuSK myasthenia gravis with and without general immunosuppression. Frontiers in Immunology, 15, 1325171.
Other organizations :
Maastricht University, MRC Weatherall Institute of Molecular Medicine, University of Oxford, University of Pisa, Medical University of Vienna, Pasteur Hellenic Institute, National and Kapodistrian University of Athens
Coating of ELISA plates with 1 µg/mL MuSK extracellular domain (ECD) produced in mammalian cells
Detection of bound MuSK antibodies with mouse anti-human IgG4:HRP (1:3,500)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
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