血管内皮抑素的原核表达及多克隆抗体的制备  

Prokaryotic Expression of Endostat in and Preparation of Polyclonal Antibody

作　　者：张一鸣[1] 王若宁[1] 李艳利[1] 顾冬民[1] 程志祥[1] 熊俊[1] 德伟[1] 陈丙莺[1] 

机构地区：[1]南京医科大学生化与分子生物学系,江苏南京210029

出　　处：《癌症》2002年第9期957-960,共4页Chinese Journal of Cancer

摘　　要：背景与目的:本实验拟在原核系统中表达并纯化人血管内皮抑素(endostatin),制备鼠抗人血管内皮抑素多克隆抗体。方法:设计引物扩增内皮抑素的cDNA,重组入原核表达载体并在大肠杆菌中诱导表达,应用纯化产物免疫3只小鼠。结果:构建成制备人血管内皮抑素的原核表达载体pQE-30,并转化入大肠杆菌BL21中得到表达产物,经Ni亲和层析柱纯化后进行SDS-PAGE鉴定结果为单一组分。利用纯化的人血管内皮抑素成功制备高滴度的鼠抗人血管内皮抑素多克隆抗体,并由Westernblot分析证实。结论:高纯度表达产物及多克隆抗体的制得为今后的研究提供了素材。Background &Objective:The aim of this study was to express an d purify human endostatin and to prepare polyclonal antibody of mouse anti-human endostatin.Methods:The cDNA of endostatin was amplified by PCR,then recombined into prokaryotic expression vector and transformed into Esc herichia coli BL21for expression;t he mice were immunized with purified products.Results:Prokaryotic expression vector pQE-30of human endostatin was successfu lly constructed;the expression product was gained after pQE-30was transfe rred into BL21.After purified by Ni a ffinity chromatography,the product was identified to be a single component by SD S-PAGE.Western blot analysis showe d that high titer mouse anti-human endostatin p olyclonal antibody was successfully prepared.Conclusion:Highly purified expression product and prepared polyclonal ant ibody provide the necessary materia l for further study.

关 键 词：制备 血管生成因子 血管内皮抑素 胶原ⅩⅧ 原核表达 多克隆抗体 

分 类 号：R730.5[医药卫生—肿瘤]