Establishment of large phage antibody library
- VernacularTitle:大容量人源噬菌体抗体库的构建
- Author:
Yuping CHEN
;
Guomin ZHANG
;
Yuanyuan QIAO
;
Gang WANG
;
Yufeng LIU
;
Bing HUA
;
Yan WANG
- Publication Type:Journal Article
- Keywords:
Phage display;
Antibody libraries;
Cre recombinase;
Single-chain Fv(ScFv)
- From:
Chinese Journal of Immunology
1985;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To create a large human phage antibody library from which easy to get diabody and select human antibody clones.Methods:VL and VH genes were amplified by RT-PCR from RNA which came from normal adult peripheral blood and new-born cord blood lymphocytes. VL and VH genes were reamplified to add a region of overlap in the ScFv linker to form ScFv genes in which VH genes were flanked by two non-homologous loxp sites. The ScFv genes were cloned into PDF to obtain a primary library. This primary library was used to infect bacteria Bs1365(expressing cre recombinase) with high multiplicity of infection(MOI) 200∶1. This procedure resulted in a very large phage antibody library through VL and VH recombined by the cre recombinase. Following recombination, phagemide were derived from these bacteria and used to infect bacteria not expressing cre(XL1-Blue) at MOI
