Screening of phage antibodies against HBsAg from the constructed phage display library and sequencing of the positive clones
- VernacularTitle:人源性抗HBsAg抗体Fab段噬菌体呈现文库的构建、筛选及阳性克隆核苷酸序列测定
- Author:
Hui GAO
;
Angang YANG
;
Lei GAO
;
Xiangrong REN
;
Zhouya YU
- Publication Type:Journal Article
- Keywords:
Hepatitis B antigens;
Antibodies;
Cloning, molecular;
DNA
- From:
Chinese Journal of Pathophysiology
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: Screening of phage antibodies against HBsAg from the constructed phage display library and sequencing of the positive clones.METHODS: RNAs were extracted from blood lymphocytes of a HBsAg-immunized volunteer. Then cDNAs were synthesized by RT-PCR. To construct the phage antibody display library, cDNAs of Fd fragments and ? chains of the antibodies were amplified through PCR and were inserted into vector pComb3H. Three rounds panning against coated HBsAg showed the specific enrichment of phage antibody. Positive clones were selected and sequenced.RESULTS: A phage antibody display library was constructed after amplifing of Fd fragments and ? chains of IgG1. Three Fab antibodies from the library were selected and sequenced. The result of sequencing showed that two of the three antibodies had the same Fd fragment and all of the three had the same light chain. CONCLUSION: The phage antibody display library was constructed successfully and three specific antibodies (Fab fragments) have been obtained from it.
