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.
Artigo
em Chinês
| WPRIM
| ID: wpr-356691
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Bacteriófagos
/
Virologia
/
Glicoproteínas de Membrana
/
Proteínas do Envelope Viral
/
Biblioteca de Peptídeos
/
Síndrome Respiratória Aguda Grave
/
Coronavírus Relacionado à Síndrome Respiratória Aguda Grave
/
Alergia e Imunologia
/
Glicoproteína da Espícula de Coronavírus
/
Genética
Tipo de estudo:
Estudo diagnóstico
/
Estudo de rastreamento
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2013
Tipo de documento:
Artigo
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