2001•Zhongguo bingli shengli zazhiRequires access
Screening of phage antibodies against HBsAg from the constructed phage display library and sequencing of the positive clones
Lei Gao
Open publisher page 0 citations
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 κ 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 κ 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.