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目的 用胃蛋白酶酶解抗哇巴因多克隆抗体 ,制备出F(ab) 2 片段 ,为改型抗体的研究提供依据。方法 免疫家兔制备出抗哇巴因多克隆抗体 ,并用胃蛋白酶分别在不同的酶解条件下消化抗体 ,继之用高效液相排阻色谱法 (HPSEC)对其进行分析、纯化 ,采用酶联免疫吸附法(ELISA)检测、鉴定其活性。结果 用胃蛋白酶 2mg/1 0 0mg抗哇巴因多克隆抗体 ,消化 1 8h ,反应体系的 pH3 0时 ,可获得F(ab) 2 片段。结论 用胃蛋白酶酶解抗哇巴因多克隆抗体是研制出有活性的F(ab) 2 片段的有效方法
Objective To digest F (ab) 2 fragment with pepsin and anti-ouabain polyclonal antibody, and to provide a basis for the study of the modified antibody. Methods Anti-ouabain polyclonal antibody was prepared by immunization in rabbits. Antibodies were digested with pepsin under different enzymatic conditions, followed by analysis by HPSEC and purification. Linked immunosorbent assay (ELISA) detection, identification of its activity. Results F (ab) 2 fragment was obtained by digesting pepsin 2 mg / 100 mg anti-ouabain polyclonal antibody for 18 h at pH 30. Conclusions Enzymatic digestion of anti-ouabain polyclonal antibody with pepsin is an effective method to develop an active F (ab) 2 fragment