Improving the affinity of an anti-TNF? scFv by error prone PCR and staggered extention process(StEP)
- VernacularTitle:联合应用错配PCR和交替延伸PCR法提高抗TNF?抗体亲和力
- Author:
Baoan WANG
;
Xiaosui CHEN
;
Yuanyuan QIAO
;
Jia QU
;
Yuxiao WANG
;
Yan WANG
- Publication Type:Journal Article
- Keywords:
Error prone PCR;
Staggered extention process;
Antibody affinity;
Dot ELISA
- From:
Chinese Journal of Immunology
1999;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To improve the affinity of an anti TNF? scFv.Methods:Starting from an anti TNF? scFv gene a mutant phage antibody library was generated by error prone PCR.Affinity improved clones were selected and subjected to staggered extension process to shuffle the mutated sites.Mutants with further improved affinity were selected by bio panning.Affinity was judged by dot blot ELISA and thiocyanate elusion ELISA.Results:Seven affinity improved mutants were obtained from library constructed by error prone PCR.By StEP mediated shuffling of these 7 clones and via bio panning,mutants with further improved affinity were obtained.Conclusion:Combination of error prone PCR and StEP could be used to improve the affinity of antibodies. [
