抗VSTM1-v2细胞因子人源化单链抗体文库的构建及筛选

发布时间：2018-05-19 05:16

本文选题：VSTM-v细胞因子 + 鼠源VH-CDR　；参考：《药学学报》2013年11期

【摘要】：本文将鼠源VH-CDR3库(library of murine VH-CDR3)移植到特定的人源scFv(single-chain antibody fragment)VH3-VΚ1框架构建人源化scFv文库,快速筛选抗VSTM1-v2的人源化单链抗体。以抗VSTM1-v2鼠源cDNA为模板,扩增出鼠源VH-CDR3库,随后将鼠源VH-CDR3库移植到人源scFv(VH3-VΚ1)上并通过核糖体展示富集抗VSTM1-v2人源化scFv文库,该文库序列通过与原核基因表达调控等成分(N端添加T7启动子、SD序列和PelB序列、C端添加his标签序列和T7 terminal序列)拼接,构建表达型TA载体转化大肠杆菌BL21(DE3)进行可溶性表达。最终构建了一个库容达1012的人源化单链抗体文库,完成了1 000个克隆的初步鉴定,并筛选出EC50达21.35 nmol·L 1的抗VSTM1-v2人源化scFv。该研究结果,一方面获得了具有潜在应用价值的抗-VSTM1-v2先导抗体;另一方面,为快速获得人源化抗体提供了一个新技术途径。
[Abstract]:In this paper, the mouse source VH-CDR3 Library (Library of murine VH-CDR3) is transplanted to a specific human source scFv (single-chain antibody fragment) VH3-V scFv 1 frame architecture humanized scFv library. The source scFv (VH3-V 1) was enriched by ribosome display and enriched in the scFv Library of anti VSTM1-v2 human origin. The library sequence was spliced by N terminal T7 promoter, SD sequence and PelB sequence, C terminal adding his label sequence and T7 terminal sequence. Finally, a human derived single chain antibody library of 1012 was constructed, and the preliminary identification of 1000 clones was completed, and the results of the anti VSTM1-v2 human scFv. EC50 up to 21.35 nmol. L 1 were screened out. On the one hand, the anti -VSTM1-v2 precursor antibody with potential application value was obtained; on the other hand, the rapid human origin was obtained. Antibodies provide a new way of technology.
【作者单位】：四川抗菌素工业研究所;成都生物制品研究所有限责任公司;
【基金】：国家“重大新药创制”科技重大专项资助项目(2011ZX09506-005) 中国医药集团新产品基金项目(2011SW12-1)
【分类号】：R392.1

【共引文献】