Screening of human phage single chain antibody against the recombinant surface protein of Streptococcus mutans and identification of its biological activities
10.3969/j.issn.1002-2694.2010.03.002
- VernacularTitle:变形链球菌重组表面蛋白(rA-P)人源单链抗体的筛选及其活性检测
- Author:
Fuxiang LI
;
Yali ZHANG
;
Shicai DING
;
Yifang ZHANG
;
Zehua WANG
;
Lan MA
- Publication Type:Journal Article
- Keywords:
Streptococcus mutans;
recombinant cell surface protein antigen;
phage library;
single chain variable fragment
- From:
Chinese Journal of Zoonoses
2010;(3):201-204,208
- CountryChina
- Language:Chinese
-
Abstract:
To isolate human phage single chain antibody against surface protein of Streptococcus mutans,the recombinant surface protein of S.mutans(rAP) was used to coat the immune tubes and the phage single chain antibody was prepared through pDAN5 phage antibody library after 5 rounds of panning.The eluted phage was enriched nearly 30 times.In these ways,13 positive clones were obtained and found to be able to bind with rAP in ELISA assay.Then one of the 13 positive clone phage plasmid was used to infect E.coli HB2151 to induce the expression of the non-fusion single chain antibody (ScFv) with IPTG induction.As demonstrated by SDS-PAGE,the molecular mass of this single chain antibody was proved to be 30 kDa and the amount of expression constituted to 30% of the total bacterial proteins.Apparently,the human phage single chain antibody against surface protein of S.mutans with biological activity was successfully screened.
