Construction and screening of SARS-CoV S protein-specific phage displayed antigen library / 病毒学报
Chinese Journal of Virology
; (6): 280-286, 2013.
Article
in Zh
| WPRIM
| ID: wpr-356691
Responsible library:
WPRO
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.
Full text:
1
Index:
WPRIM
Main subject:
Bacteriophages
/
Virology
/
Membrane Glycoproteins
/
Viral Envelope Proteins
/
Peptide Library
/
Severe Acute Respiratory Syndrome
/
Severe acute respiratory syndrome-related coronavirus
/
Allergy and Immunology
/
Spike Glycoprotein, Coronavirus
/
Genetics
Type of study:
Diagnostic_studies
/
Screening_studies
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Virology
Year:
2013
Type:
Article