Secretory expression and biological activity analysis of an anti-H5 single-chain antibody from Pichia pastoris / 病毒学报
Chinese Journal of Virology
;
(6): 202-206, 2011.
Artigo
em Chinês
| WPRIM
| ID: wpr-286054
ABSTRACT
In our previous study, a panel of 52 broadly cross-reactive H5-specific monoclonal antibodies (MAbs) were generated and characterized. The 13D4, one of these MAbs, has been demonstrated to protect mice against lethal challenge by 4 strains of H5N1 avian influenza virus representing the currently prevailing genetic populations, clades 1, 2.1, 2.2, and 2.3. Here, we further cloned the gene of the 13D4 MAb and constructed a single-chain variable fragment. Then, the 13D4 single-chain antibody (scFv) was expressed in secretory maner in Pichia pastoris. The supernatant of the culture was concentrated and subjected to ammonium sulfate precipitation. The purity of the 13D4 scFv was around 90% in SDS-PAGE following ion-exchange chromatography. We further investigated its binding property using hemagglutination inhibition (HI) test and blocking ELISA. The results indicated that the 13D4 scFv shared the same binding sites and comparable HI titer with the prototype murine 13D4 Mab. In conclusion, an anti-H5 single-chain wide-spectrum neutralizing antibody is prepared successfully in yeast system.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Pichia
/
Testes de Inibição da Hemaglutinação
/
Fragmentos de Imunoglobulinas
/
Alergia e Imunologia
/
Virus da Influenza A Subtipo H5N1
/
Anticorpos de Cadeia Única
/
Genética
/
Anticorpos Antivirais
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2011
Tipo de documento:
Artigo
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