Characterization of a monoclonal antibody specific for the HIV-1 precursor glycoprotein.
HIV-1 infected Molt4-T4 cells provide an efficient system for the production of cellular precursor gp160 of HIV envelope glycoproteins, gp120 and gp41. The precursor gp160 was purified on an immuno-affinity column containing antibodies from sera of HIV-1-seropositive patients. The precursor gp160 was then isolated by preparative polyacrylamide gel electrophoresis. Two out of four Balb/c mice, immunized with these purified preparations of gp160, developed specific circulating antibodies. A hybridoma cell line was subsequently isolated producing monoclonal antibody KL49/19 (IgG1, K) specific for gp160. This monoclonal antibody can specifically immunoprecipitate gp160, existing in HIV-1-infected cells. In an immunoblotting assay, it identifies mainly gp160 and shows a slight affinity for the mature glycoprotein, gp120. The monoclonal antibody is probably directed against an epitope in the polypeptide residue of gp160 since it can recognize a deglycosylated polypeptide of molecular weight 90,000, a product of gp160 digestion by endoglycosidase H (Endo H). It does not cross-react with any protein of HIV-2 by immunoblot or immunoprecipitation assays. By virtue of its specificity, the monoclonal antibody KL49/19 might provide a powerful probe with which to detect gp160 in cells which might partially express the HIV-1 genes.
Krust
B
B
Unité d'Oncologie Virale, Institut Pasteur, Paris, France.
Laurent
A G
AG
Le Guern
A
A
Jeannequin
O
O
Montagnier
L
L
Hovanessian
A G
AG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity; D002460:Cell Line; D006678:HIV / Q000276:immunology*; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D006801:Humans; D011498:Protein Precursors / Q000276:immunology; D012191:Retroviridae Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D011498:Protein Precursors; D012191:Retroviridae Proteins