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肝癌特异性鼠源及人源化单链抗体基因的构建及大肠杆菌中的表达
引用本文:袁清安,俞炜源.肝癌特异性鼠源及人源化单链抗体基因的构建及大肠杆菌中的表达[J].生物工程学报,2000,16(1):86-90.
作者姓名:袁清安  俞炜源
作者单位:军事医学科学院生物工程研究所,北京
摘    要:为探讨一株肝细胞癌特异性鼠源及其人源化单链抗体基因在大肠杆菌中的可溶性表达策略并比较二者对抗原的结构能力,在三种载体中分别以融合、分泌及胞内表达的方式进行了研究,表达产物均以包涵体形式存在;对复性后的单链抗体以细胞ELISA及竞争抑制流式细胞仪法进行检测,表明人源化单链抗体和鼠源单链抗体有相近的抗原结合能力。结论是:大肠杆菌中表达的基因工程单链抗体的可溶性可能主要由自身氨基酸一级序列决定;先前的设

关 键 词:单链抗体  肝癌  大肠杆菌  免疫原性  可溶性表达
文章编号:1000-2061(2000)01-0086-05
修稿时间:1998-06-08

Construction and Expression of A Hepatocellular Carcinoma Specific Rodent and Its Humanized Single-chain Fv Fragments in Escherichia coli
YUAN Qing-an ,YU Wei-yuan ,HUANG Cui-fen.Construction and Expression of A Hepatocellular Carcinoma Specific Rodent and Its Humanized Single-chain Fv Fragments in Escherichia coli[J].Chinese Journal of Biotechnology,2000,16(1):86-90.
Authors:YUAN Qing-an  YU Wei-yuan  HUANG Cui-fen
Institution:YUAN Qing-an ,YU Wei-yuan ,HUANG Cui-fen ;(Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071)
Abstract:The aim of this research is to exploit the expression strategies for two genes encoding a rodent and its humanizod version single-chain fragments(scFv) specific for hepatocellular carcinoma (HCC) and compare these two scFves in antigen binding activity. Three vectors were used to express these two genes in different riytes of fusion pathway, secrete pathway and intmcellular pathway. The vefolded single-chain antibodies were examined by antigen capture ELISA. Results showed that inclusion bodies were produced in all of the applied vectors despite of variation of IPTG concentration and culture temperature. Cell ELISA and binding competition inhibition indicated that the humanized single-chain Fv retained the similar affinity as its rodent counterpart. These results implied that the solubility of genetic antibodies are determined primarily by its amino acids sequence; The adopted humnanization strategy in previous design made little effect on the natural conformation of complementarity-determining regions(CDR)of the parent antibody. The humanized HCC specific scFv can be further evaluated and developed as therapeutics.
Keywords:Humanization  single-chain antibody  hepatocellular carcinoma  expression
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