Dwyer D S, Bradley R J, Urquhart C K, Kearney J F
J Immunol Methods. 1983 Feb 25;57(1-3):111-9. doi: 10.1016/0022-1759(83)90069-8.
An enzyme-linked immunoabsorbent assay has been developed for measuring anti-acetylcholine receptor antibodies from sera of patients with myasthenia gravis or tissue culture supernatants from hybridomas. Acetylcholine receptor from a detergent extract of muscle tissue was bound indirectly to microtiter plates via a monoclonal anti-receptor antibody already coupled to the polyvinyl plates. Myasthenic sera or antibodies in tissue culture media were then tested for binding to the acetylcholine receptor attached to the monoclonal antibody. Anti-receptor antibodies were detected in the serum of 80% of myasthenic patients when assayed by this method and the levels of antibody corresponded fairly well with antibody titers determined by an immunoprecipitation assay. Occasional myasthenic patients had serum antibodies which reacted specifically with the monoclonal antibody attached to the microtiter plate. The assay described here was far less time-consuming than immunoprecipitation assays, required only small quantities of receptor, and did not require the use of radioisotopes such as 125I-alpha-bungarotoxin.
已开发出一种酶联免疫吸附测定法，用于检测重症肌无力患者血清中的抗乙酰胆碱受体抗体或杂交瘤的组织培养上清液中的该抗体。来自肌肉组织去污剂提取物的乙酰胆碱受体通过已偶联到聚乙烯板上的单克隆抗受体抗体间接结合到微量滴定板上。然后检测重症肌无力患者血清或组织培养基中的抗体与附着在单克隆抗体上的乙酰胆碱受体的结合情况。用该方法检测时，80%的重症肌无力患者血清中可检测到抗受体抗体，且抗体水平与免疫沉淀法测定的抗体滴度相当吻合。偶尔有重症肌无力患者的血清抗体可与附着在微量滴定板上的单克隆抗体发生特异性反应。此处描述的测定法比免疫沉淀法耗时少得多，只需要少量受体，且无需使用如125I-α-银环蛇毒素等放射性同位素。
