Construction and Verification of High-capacity Ribosome Display Single-chain Fv Library
- VernacularTitle:大容量核糖体展示抗体库的构建与初步鉴定
- Author:
Kaihong HUANG
;
Xuexian LI
;
Yingting CHEN
- Publication Type:Journal Article
- Keywords:
Ribosomes;
Single-chain Fv library;
Gastrointestinal carcinoma
- From:
Chinese Journal of Practical Internal Medicine
2006;0(18):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct high-capacity ribosome display single-chain Fv library for selection of high affinity ScFv antibody.Methods We isolate human lymphocyte from peripheralblood(2 normal,3 gastric cancer,3 colonic cancer,1 pancreatic cancer,each 5 mL and 2 newborn,each 2 mL)and extract RNA for cloning whole human heavy chain and light chain gene by RT-PCR.VH and VL were rearranged randomly by SOEing(splicing by overlap extension,SOEing).Finally,the elements for in vitro screening such as T7 promoter and ribosome binding site were introduced while the SOEing products were amplified.Moreover,ribosome display template were verified by blue/white screening and further sequencing.Results We successfully constructed ribosome display ScFv library with a volume of 1.1?1013.Conclusion The construction of high-capacity ScFv library shed light on multiple therapeutic ScFv screening.
