Large-scale production and purification of a vaccinia recombinant-derived HIV-1 gp160 and analysis of its immunogenicity.
AIDS Res Hum Retroviruses
; 5(2): 159-71, 1989 Apr.
Article
em En
| MEDLINE
| ID: mdl-2713166
ABSTRACT
The human immunodeficiency virus (HIV-1) envelope gene was expressed in large-scale microcarrier cultures of Vero cells using a system involving coinfection with two recombinant vaccinia viruses. One recombinant contained the bacteriophage T7 RNA polymerase gene under control of a vaccinia virus promoter. The second contained the HIV-1 gp160 gene flanked by T7 promoter and termination sequences. The protein was expressed on the surface of infected cells, and it was shown to have a molecular weight of 160 kD and to react with gp41 and gp120 specific monoclonal antibodies. After purification by successive affinity and ion-exchange chromatography, the protein was demonstrated to be present in a particulate form with a diameter in the range of 15-30 nm. When injected into goats a high-titer gp160 specific antibody response was elicited and group-specific neutralizing activity could be demonstrated in vitro. The immunogenicity of the protein was also studied in conjunction with a number of adjuvant formulations, and the highest potency in mice was obtained using a preparation with 0.2% Al(OH)3 and 0.25% deoxycholate.
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Base de dados:
MEDLINE
Assunto principal:
Vacinas
/
Vacinas Sintéticas
/
Proteínas dos Retroviridae
/
Proteínas do Envelope Viral
/
HIV-1
Limite:
Animals
Idioma:
En
Ano de publicação:
1989
Tipo de documento:
Article