| 基因工程技术制备人源抗HBsAg抗体Fab片段 |
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| 引用本文： | 尤长宣,罗荣城,丁雪梅,韩焕兴,胡栋平,范列英.基因工程技术制备人源抗HBsAg抗体Fab片段[J].现代免疫学,2000,20(1):33-34,37. |
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| 作者姓名： | 尤长宣 罗荣城 丁雪梅 韩焕兴 胡栋平 范列英 |
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| 作者单位： | 1. 第一军医大学南方医院肿瘤科,广州,510515
2. 第二军医大学长征医院全军临床免疫中心,上海200003 |
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| 基金项目： | 本项目受国家自然科学基金和广州市重点科研项目基金资助(项目号39670668、97-Z-65-02) |
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| 摘 要： | 将从抗体文库中筛选出的抗HBsAg Fab阳性克隆转化大肠肝杆菌 ,IPTG诱导表达 ,经Western blot鉴定 ,证实上清中含有可溶性抗HBsAg Fab片段 ,应用自制的羊抗人免疫球蛋白片段抗体 (anti Fab )与Gama bind交联制备的亲和层析柱纯化表达产物 ,所得纯化产物在SDS PAGE中形成单一条带 ,经Western blot证实具备良好抗原特异性 ,Dot blot证实具备良好抗原结合活性。
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| 关 键 词： | HBsAg 抗体 Fab 亲和层析 |
| 文章编号： | 1001-2478(2000)01-0033-02 |
Preparation of Human Anti-HBsAg Fab Fragment by Genetic Engineering Technology |
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| You Changxuan,Luo Roncheng,Ding Xuemei,Han Huanxing,Hu Dongping,Fan Lieying.Preparation of Human Anti-HBsAg Fab Fragment by Genetic Engineering Technology[J].Current Immunology,2000,20(1):33-34,37. |
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| Authors: | You Changxuan Luo Roncheng Ding Xuemei Han Huanxing Hu Dongping Fan Lieying |
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| Institution: | You Changxuan
,Luo Roncheng
,Ding Xuemei
;(The Cncology Department, Nanfang Hospital, Guangzhou,510515);Han Huanxing
,Hu Dongping
,Fan Lieying
;( Clinical Immunology Center of PLA, Changzheng Hospital,Shanghai, 200003) |
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| Abstract: | The specific anti HBsAg clone was selected from the established combinatorial library and transformed into E coli xL1 blue by using IPTG as inducing agent for the expression of Fab fragment As verified by Western blot,soluble anti HBsAg Fab fragment could be detected in the supernatants of cells by repeated freezing and thawing The expressed products were purified by affinity chromatography column prepared with sheep anti human Fab IgG and Gama bind Sepharose The purified Fab fragment formed a single band in SDS PAGE,and possessed good antigenic specificity as well as the excellant combining activities with antigens as verified by Dot blot |
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| Keywords: | HBsAg antibody Fab affinity ehromatographye |
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