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本文根据独特型-抗独特型理论探讨Graves′病的发病机理,报道了对兔抗hTSHIgG免疫原的分离,纯化及鉴定。首先用饱和硫酸铵粗提兔抗hTSH血清γ-球蛋白,继以葡萄球菌A蛋白琼脂糖凝胶亲和层析柱(ProteinAsepharoseCL4B)纯化IgG,经十二烷基硫酸钠—聚丙烯凝胶电泳(SPS-PAGE)鉴定,证实为IgG纯品。再经TSH偶联的亲和层析柱为免疫吸附剂,进一步分离出抗TSHIgG,然后用放射免疫分析(RIA)检测证实保持了较高的抗体活性。
In this paper, based on the idiotype-anti-idiotype theory to explore the pathogenesis of Graves’ disease, the isolation, purification and identification of rabbit anti-hTSHIgG immunogen was reported. Rabbit anti-hTSH serum γ-globulin was first roughly extracted with saturated ammonium sulfate and then IgG was purified by staphylococcal Protein A Sepharose CL4B. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SPS-PAGE) identification, confirmed as pure IgG. The TSH-conjugated affinity chromatography column was used as the immunoadsorbent to further isolate TSHIgG, and then the high antibody activity was confirmed by radioimmunoassay (RIA).