A set of mAbs for binding to MuSK, AChR, and MOG were used as controls. Cell culture supernatant from either an established murine MuSK mAb (4A3; ref. 32 (link)) or a murine MOG mAb (8-18C5; ref. 64 (link)) hybridoma was applied to a Protein G/Sepharose column (GE Healthcare) to isolate the IgG. We also engineered the MuSK mAb (4A3) and the MOG mAb (8-18C5) as chimeric mouse-human recombinant mAbs. They were produced to contain the murine mAb heavy and light chain variable regions fused to the respective human constant regions using an approach we described (65 (link)). These chimeric recombinant mAbs served as positive controls in the human antibody–binding assays and did not require a substitute (murine-specific) secondary antibody because the constant regions were identical to those of human mAbs. The AChR mAb (clone 637) was derived from a human MG thymus (24 (link), 25 (link), 66 (link)). The variable regions were synthesized as gBlock gene fragments (Integrated DNA Technologies), then subcloned into expression vectors, expressed, and purified using approaches we described (67 (link)).
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