Epitope mapping of a monoclonal antibody against the Gp85 of avian leukosis virus subgroup J.
J Vet Med Sci
; 74(6): 693-7, 2012 Jun.
Article
em En
| MEDLINE
| ID: mdl-22214862
ABSTRACT
Avian leukosis virus subgroup J poses a great threat to the poultry industry in China. To reduce the economic losses, a quick method for detection of ALV-J antigen is required for diagnosis and identification of the congenitally transmitting hens. In this study, we report the production and evaluation of one monoclonal antibody (MAb) suitable for achieving these goals. The gp85 gene of avian leukosis virus subgroup J CAUHM01 China isolates was subcloned into the expression vectors pGEX-6p-1 and pET28a and successfully expressed in E. coli. After immunizing BALB/c mice with recombinant His-Jgp85 protein, splenic cells from immunized mice were fused with SP2/0 myeloma cells to produce hybridomas. We isolated and characterized one ALV-J gp85-specific MAb by determining its titer, affinity and IgG subclass. In addition, we performed epitope mapping and determined the epitope for the MAb 1E3 to be 81-92 aa of ALV-J gp85 protein (LPWDPQELDILG). Bioinformatics analysis and IFA studies revealed that this epitope is conserved among all ALV-J isolates and that this antibody could serve as a useful reagent for ALV-J detection and diagnosis.
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Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Leucose Aviária
/
Proteínas do Envelope Viral
/
Vírus da Leucose Aviária
/
Mapeamento de Epitopos
/
Anticorpos Monoclonais
/
Antígenos Virais
Tipo de estudo:
Prognostic_studies
Limite:
Animals
País/Região como assunto:
Asia
Idioma:
En
Revista:
J Vet Med Sci
Ano de publicação:
2012
Tipo de documento:
Article