【摘要】 目的制备哇巴因多克隆抗体F(ab) 2 片段 ,并对其进行分子量的测定。方法免疫家兔制备哇巴因多克隆抗体 ,在适宜的条件下 ,用胃蛋白酶酶解哇巴因多克隆抗体 ,制备F(ab) 2 片段。酶解产物经HPSEC分析、纯化 ,ELISA法检测其免疫学活性。并在适宜的色谱条件下 ,分别测定三种标准蛋白的洗脱体积 ,绘制标准曲线 ,测定其分子量。结果 10 0mg哇巴因多克隆抗体在 2mg胃蛋白酶消化 18h ,反应体系pH 3 .0的条件下消化 ,制备出了活性的F(ab) 2 片段 ,其相对分子质量为 10 7kD。结论用胃蛋白酶消化哇巴因多克隆抗体 ,可制备出活性的F(ab) 2 片段 ,同时其相对分子质量的测定为其进一步研究提供了依据。更多还原

【Abstract】 PurposeThe aim is to prepare ouabain polyclonal antibody F(ab) 2 fragment and to estimate its molecular weight.MethodsOuabain polyclonal antibody was obtained from immunized rabbits.The antibody was digested with pepsin.The resulting products were analyzed and the molecular weight of F(ab) 2 fragment was estimated by HPSEC.The immune activity was detected by ELISA.Results100 mg of ouabain polyclonal antibody was digested by 2 mg of pepsin for 18 hours at pH 3.0 and active ouabain polyclonal antibody F(ab) 2 framgment was obtained.Its molecular weight was 107 kD.ConclusionThe active ouabain polyclonal antibody F(ab) 2 fragment could be prepared by digesting its antibody with pepsin.更多还原