RESUMO
In a previous work we selected montanide ISA720 (M-ISA720) among different adjuvants for the vaccination with a V3 loop based multi-epitope polypeptide TAB9. In this paper we present the evaluation of the toxicity and immunogenicity of this formulation in non-human primates. TAB9 in M-ISA720 was highly immunogenic in macaques (Macaca fascicularis) inducing antibodies against TAB9 in all animals after one inoculation and a strong anamnestic response after booster injections. Furthermore 97% of the V3 peptides included were recognized by TAB9 sera. No differences between doses of 200 microg and 1 mg of TAB9 in M-ISA720 were observed after four immunizations. Neutralizing antibodies against five HIV-1 isolates were detected in most animals. Animals remain healthy throughout the study and did not show lesions at the inoculation site.
Assuntos
Adjuvantes Imunológicos/administração & dosagem , Anticorpos Antivirais/biossíntese , Infecções por HIV/imunologia , HIV-1/imunologia , Epitopos Imunodominantes/imunologia , Manitol/análogos & derivados , Ácidos Oleicos/imunologia , Peptídeos/imunologia , Sequência de Aminoácidos , Animais , Afinidade de Anticorpos , Sequência Consenso , Ensaio de Imunoadsorção Enzimática , Macaca radiata , Manitol/imunologia , Dados de Sequência Molecular , Testes de Neutralização , ÓleosRESUMO
Recombinant hepatitis B surface antigen (r-HBsAg) produced in yeast is adsorbed on a diatomaceous earth matrix for purification purposes. A pH dependence in the adsorption-elution behavior was found. The capacity of celite (Hyflo Super Cei) for adsorbing r-HBsAg increased with decreasing pH. Nonspecific proteins were also adsorbed, but a low pH dependence was found. Elution from the matrix was performed using a basic pH buffer, in which r-HBsAg is more specifically adsorbed/desorbed than contaminant proteins, permitting the purification of the r-HBsAg. A pH of 4.0 was used for adsorption and pH 8.2 was used for desorption. The described protocol allows a purification factor between three- and fivefold with respect to contaminant proteins and sixfold with respect to contaminant DNA. Finally, the adsorption step was successfully scaled-up for production purposes.