title: The stability of solubilized mammalian muscle acetylcholine receptors during purification by monoclonal immunoadsorption.
abstract: The stability of nicotinic acetylcholine receptors (AChR) solubilized from mammalian skeletal muscle in nonionic detergent was investigated under various conditions of pH, chaotropic ions, and unfolding reagents in order to allow its purification in high yield by immunoadsorption to monoclonal antibodies. Preservation of the antigenicity and/or binding sites for alpha-bungarotoxin was used as an indicator of the receptor protein's integrity. Both were preserved in the pH range 6.5-8.0, but when exposed for 1 h at 4 degrees C to a pH outside this range, greater than 50% activity was lost. Of the chaotropic ions studied (NaSCN, NaI, NaNO3, NaCl), only NaCl was tolerated. Most of the AChR's toxin-binding activity was preserved after exposure to 2 M NaCl, which was suitable for dissociating AChR when a monoclonal antibody with relatively low binding affinity was selected as the immunoadsorbent. Yields of purified AChR were optimal (30%) when a low amount of monoclonal antibody was coupled to cyanogen bromide-activated agarose (1 mg protein/ml gel).
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D005260:Female; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D009132:Muscles / Q000032:analysis*; D011950:Receptors, Cholinergic / Q000187:drug effects / Q000302:isolation & purification*; D012995:Solubility; D014101:Torpedo; D014508:Urea / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D006146:Guanidines; D011950:Receptors, Cholinergic; D014508:Urea; D019791:Guanidine