摘要
【目的】构建前列腺癌患者的噬菌体抗体库,为前列腺癌特异性的人源性抗体Fab片段的筛选及其应用奠定基础。【方法】由前列腺癌患者外周血分离得淋巴细胞,完整提取其总RNA,经RT-PCR及半套式扩增得到免疫球蛋白全部轻链和重链Fd段基因,然后经酶切、纯化、连接等步骤克隆进载体pComb3,并电转化大肠杆菌XL1-Blue,从而构建前列腺癌患者噬菌体抗体库。【结果】成功构建噬菌体抗体Fab片段库,其轻链及重链Fd段与载体DNA的重组率分别为87%、79%,库容为23.2×107,滴度为87×1012mL-1。【结论】本研究成功构建了前列腺癌患者噬菌体抗体库。
To construct the Fab phage antibody library of prostatic cancer patients for selection of human specific antibodies against prostate cancer and their future application. Lymphocytes were isolated from peripheral blood of prostatic cancer patients.tRNA was isolated and the light chains and heavy chain Fd fragments of immunoglobulin gene were amplified by PCR and semi nest PCR,which products were cloned into vector pComb3 and then translocated into E.Coli XL1 Blue. The immune Fab phage antibody library of prostatic neoplasm was constructed.The recombination rates of light chains and Fd fragments were 87%and 79%, respectively.The volume of the phage library was 23.2×106 and the titre was 87×1012 mL-1.[Conclusion]The phage antibody library of prostatic cancer patients is constructed successfully.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2004年第4期299-303,共5页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省自然科学基金资助项目(001356)
关键词
前列腺癌
噬菌体
抗体库
肿瘤
prostatic neoplasms
bacteriophages
antibodies
