Brett Einarson

Production and assay of antibodies to acetylcholine receptors.

Publisher Summary This chapter describes the production and assay of antibodies to acetylcholine receptors. All the basic methods necessary to assay and purify acetylcholine receptors (AChRs) and AChR subunits for use as immunogens to produce antisera and monoclonal antibodies (mAbs) to AChRs and to induce experimental autoimmune myasthenia gravis are discussed in the chapter. AChR is routinely identified and quantitated, by the binding of radioactively labeled krait or cobra venom toxins, which are competitive inhibitors of acetylcholine binding. An easy approach to study the specificity of antibodies to intact AChR is to test their reaction, with 123 I-toxin-labeled AChR, from various species by radioimmunoassay. The competitive binding to native AChR method uses mAbs to AChR that have been mapped, by their ability to react, with denatured subunits to help map the subunit specificity of antibodies, that react only, with native AChR, and to distinguish between mAbs to various antigenic determinants on the same subunit. It is demonstrated that the antigenicity of the native AChR molecule is dominated, by a small region, on the α-subunit.