Construction and screening of SARS-CoV S protein-specific phage displayed antigen library.
- Author:
Rui-Ping WU
1
;
Jia-Zi MENG
;
Yu-Xian HE
Author Information
1. Institute of Medical Virology, School of Laboratory Medicine, Wenzhou Medical College, Wenzhou 325035, China. wuruiping_q@126.com
- Publication Type:Journal Article
- MeSH:
Antibodies, Viral;
immunology;
Bacteriophages;
genetics;
metabolism;
Epitopes;
genetics;
immunology;
Humans;
Membrane Glycoproteins;
genetics;
immunology;
Peptide Library;
SARS Virus;
genetics;
immunology;
Severe Acute Respiratory Syndrome;
immunology;
virology;
Spike Glycoprotein, Coronavirus;
Viral Envelope Proteins;
genetics;
immunology
- From:
Chinese Journal of Virology
2013;29(3):280-286
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study is to construct a SARS-CoV S protein-specific phage displayed antigen library for the epitope characterization of anti-S monoclonal antibodies (mAbs). First, the full-length gene of SARS-S protein was PCR amplified, purified and then digested with DNase I to obtain DNA fragments in the size range of 50-500 bp. The resulting fragments were blunt-end ligated to the modified phage display vector pComb3XSS. The reactions were electrotransformed into XL1-Blue and infected with VCSM13 helper phage. The SARS-CoV S protein-specific phage displayed antigen library was biopanned and screened against two anti-S mAbs, S-M1 and S-M2. The results showed that we successfully constructed the phage displayed antigen library with a size of 5.7 x 10(6). After three-rounds of biopanning, 14 positive phage clones for S-M1 and 15 for S-M2 were respectively identified. Sequence analyses revealed the possible epitopes of two mAbs. Therefore, the S protein-specific phage displayed antigen library provides a crucial platform for the epitope characterization of anti-S antibodies and it is highly valuable for development of SARS vaccines and diagnostics.
