| 抗人VEGF165单链抗体的构建与表达 |
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| 引用本文： | 赵泽国,杨治华,冉宇靓,刘军,孙立新,董志伟.抗人VEGF165单链抗体的构建与表达[J].免疫学杂志,2000,16(2):92-95. |
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| 作者姓名： | 赵泽国 杨治华 冉宇靓 刘军 孙立新 董志伟 |
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| 作者单位： | 中国医学科学院,中国协和医科大学肿瘤研究所,北京,100021 |
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| 基金项目： | 国家863计划资助项目(863-102-09-02-03) |
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| 摘 要： | 为降低鼠源抗体对人体的免疫原性，构建表达了ＶｍＤ１１的单链抗体。方法用ｏｖｅｒｌａｐＰＣＲ构建出单链抗体的基因，克隆入表达载体ｐＫｐＬ－３ａ，在大直菌ｏｐｏ２１３６中表达，采用抗原结合ＥＬＩＳＡ和竞争抑制ＥＬＩＳＡ测定活性。结果经ＳＤＳ－ＰＡＧＥ检测，诱导后的细菌表达了２６ｋＤ的蛋白；ＷｅｓｔｅｒｎＢｌｏｔ证实该蛋白为表达的单链抗体。经变性，复性后，该单链抗体可特异结合人ＶＥＧＦ１６５抗原，并与Ｖ
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| 关 键 词： | 人VEGF165 单链抗体 原核表达 肿瘤 |
| 文章编号： | 1000—8861(2000)02,0092—04 |
| 修稿时间： | 1999－06-17 |
Construction and expression of anti--human VEGF165 ScFv |
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| ZHAO Ze-guo
,YANG Zhi-hua
,RAN Yu-liang
,LIU Jun
,SUN Li-xin
,DONG Zhi wei.Construction and expression of anti--human VEGF165 ScFv[J].Immunological Journal,2000,16(2):92-95. |
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| Authors: | ZHAO Ze-guo YANG Zhi-hua RAN Yu-liang LIU Jun SUN Li-xin DONG Zhi wei |
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| Institution: | ZHAO Ze-guo
,YANG Zhi-hua
,RAN Yu-liang
,LIU Jun
,SUN Li-xin
,DONG Zhi wei
;(Cancer Institute, Chinese Academy of Medical Science, Peking Union Medical College, Beijing 100021,China) |
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| Abstract: | Objective In order to reduce the immunogenicity of murine antibody.a ScFv against human VEGF165 was expressed in E-cole. Methods The variable region genes of heavy chain and light chain were amplilied by PCR. The ScFv gene was produced by overlap PCR of VH and VL,then ligated into expression vector pKpL-3a,and expressed in E.coli pop2136.The antigen binding activity of the ScFv was assayed by antigen binding ELISA and competition ELISA with the parent McAb. Resulls With the metbod of SDS-PAGE, western blot,antigen binding ELISA and competilion ELISA,it confirmed that E coli, expressed the correct 26kD of ScFv,which could specifically bind to the huaman VEGF165 and recognized the same epitope with the parent McAb. Conclusion An anti-human VEGI165 ScFV was successfully constructed expressed in E-coli. |
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| Keywords: | human VEGF155 ScYv procaryotic expression |
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