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用EBV转化的大肠癌患者B细胞构建噬菌体抗体库 被引量:4

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摘要 目的 构建全人源化的噬菌体抗体库。方法 采用EBV转化技术,ELISA技术,PCR技术,噬菌体表面呈现技术。结果 EBV转化8例大肠癌患者PBMC,其中七例有抗大肠癌抗体产生,多次PCR扩增出VH(γμ)和9种VL(κ,λ)基因,经简并组合成15种ScFv基因。在体外与表达载体fUSE5连接,电导入大肠杆菌MC1061,经四环素抗性筛选,得到库容为1.1×10^7的初级噬菌体抗库。全长SvFv基因的插入库为70%。结论 应用EBV转化技术联菌体抗体库技术,制备全人源化的抗肿瘤单链抗体(SvFv),这一策略是完全可行的。
出处 《细胞与分子免疫学杂志》 CAS CSCD 2000年第6期495-498,共4页 Chinese Journal of Cellular and Molecular Immunology
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