B细胞淋巴瘤Fab噬菌体抗体库的构建  

Construction of Fab Fragments Phage Display Library for Human B-lymphoma

作　　者：申咏梅[1] 杨晓春[1] 董宁征[2] 谢小芳[1] 白霞[2] 

机构地区：[1]苏州大学附属第二医院,江苏苏州215004 [2]江苏省血液研究所苏州大学附属第一医院,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2006年第4期589-592,共4页Suzhou University Journal of Medical Science

基　　金：国家自然科学基金资助课题(30400111);江苏省自然科学基金资助课题(BK2004041)

摘　　要：目的构建B细胞淋巴瘤Fab噬菌体抗体库。方法从人B细胞淋巴瘤细胞系Raji细胞免疫的BALB/c小鼠脾细胞中提取总RNA,RT-PCR扩增免疫球蛋白轻链κ基因及重链Fd片段,然后经酶切、纯化、连接等步骤将轻链κ基因和重链Fd片段依次克隆入噬菌体载体pComb3H-SS中,并电转化大肠杆菌XL1-Blue。结果构建了抗人B细胞淋巴瘤Fab噬菌体抗体库,轻链κ基因、重链Fd片段与载体的重组率分别为100%和78%,库容量为2.18×107。结论成功构建了抗人B细胞淋巴瘤Fab噬菌体抗体库,方便进一步筛选抗人B细胞淋巴瘤抗体。Objective To construct Fab phage display library against human B-lymphoma. Methods The total RNA was extracted from the spleen cells of the immunized BALB/c mice with Raji B-lymphoma cell strain and the immunoglobulin genes of the light chain k and the heavy chain Fd fragments were amplified by RT-PCR. After restrictive digestion, purification and ligation, the light chain and the heavy chain Fd were subsequently inserted into the phagmid vector pComb3H-SS, and then eleetroprated into E. coli XL1-Blue. Results The Fab phage display library against B-lymphoma was constructed. The combination rate of the light chains and heavy chain Fd fragments was 100 % and 78 % respectively. The volume of Fab phage library was reached 2.18 × 107. Conclusion The Fab phage display library against human B-lymphoma is constructed successfully. The present study can be used as foundation for succeeding screening of specific antibody against human B-lymphoma.

关 键 词：B细胞淋巴瘤 FAB 噬菌体抗体库 

分 类 号：R392.11[医药卫生—免疫学]