| 哇巴因多克隆抗体F(ab)_2片段的制备及其分子量测定 |
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| 引用本文： | 张明娟,吕卓人,袁育康,贺浪冲,王颢.哇巴因多克隆抗体F(ab)_2片段的制备及其分子量测定[J].中国生化药物杂志,2001(1). |
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| 作者姓名： | 张明娟 吕卓人 袁育康 贺浪冲 王颢 |
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| 作者单位： | 西安交通大学第一医院心内科!陕西西安710061(张明娟，吕卓人，王颢)，西安交通大学第一医院医学院免疫教研室!陕西西安710061(袁育康)，西安交通大学第一医院药学院仪器分析中心!陕西西安710061(贺浪冲) |
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| 基金项目： | 国家科技部医药技术创新博士项目!(98 B 0 7) |
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| 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。
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| 关 键 词： | 哇巴因 抗体 F(ab)_2片段 制备 相对分子量 |
Preparation and molecular weight estimation of ouabain polyclonal antibody F(ab)_2 fragment |
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| ZHANG Ming juan ,LU Zhuo ren ,YUAN Yu kang ,HE Lang chong ,WANG Hao.Preparation and molecular weight estimation of ouabain polyclonal antibody F(ab)_2 fragment[J].Chinese Journal of Biochemical Pharmaceutics,2001(1). |
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| Authors: | ZHANG Ming juan LU Zhuo ren YUAN Yu kang HE Lang chong WANG Hao |
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| Institution: | ZHANG Ming juan 1,LU Zhuo ren 1,YUAN Yu kang 2,HE Lang chong 3,WANG Hao 1 |
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| 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. |
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| Keywords: | Ouabain Antibody F(ab) 2 fragment Preparation Molecular weight |
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