抗人大肠癌抗体CAb1 Fab基因的克隆、表达和鉴定
结直肠肿瘤,,结直肠肿瘤;,Fab抗体;克隆;表达,抗人大肠癌抗体CAb1Fab基因的克隆、表达和鉴定,0引言,1材料和方法,2结果,3讨论,【参考文献】
Identification, expression and cloning of Fab gene of MAb CAb1 against human colorectal cancerYANG XiangMin, XING JinLiang, YAO XiYing, ZHANG SiHe, CHEN ZhiNan
Cell Engineering Research Center, Fourth Military Medical University, Xi’an 710033, China
【Abstract】 AIM: To identify and express the Fab Fragment against human colorectal cancer in E.coli by cloning the Fab genes of the mAb CAb1. METHODS: Light chain and Fd fragments of MAb CAb1 were amplified by RTPCR and PCR products were sequenced and analyzed after being cloned into pMD18T vector. The expression vector FdL/pComb3 was obtained after subcloning light chain and Fd fragment gene into pComb3 and deleting the gⅢ gene. Then the vector FdL/pComb3 was transformed into XL1Blue E.coli strain and induced by IPTG. The specificity of the expression products was detected by ELISA and immunofluorescence staining methods. RESULTS: The Fab gene of MAb CAb1 was cloned and expressed in E.coli. The expression product CAb1 Fab possessed the activity to combine with cell strain SW480. CONCLUSION: We successfully construct an expression vector FdL/pComb3 to express monovalent Fab against human colorectal cancer. ......
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