Partial purification of native HIV transmembrane protein gp41: generation of polyclonal and monoclonal antibodies

• Published in: AIDS research and human retroviruses Vol. 4; no. 1; p. 51
• Main Authors: Zweig, M, Showalter, S D, Bladen, S V, Gilden, R V, Arthur, L O, Robey, W G, Nara, P L, Fischinger, P J
• Format: Journal Article
• Language: English
• Published: United States 01.02.1988
• Subjects: Antibodies, Monoclonal - immunology; Antibodies, Viral - immunology; Cell Line; Chromatography, Affinity; HIV - analysis; HIV - immunology; HIV Antibodies; HIV Envelope Protein gp41; Humans; Neutralization Tests; Retroviridae Proteins - immunology; Retroviridae Proteins - isolation & purification; Viral Envelope Proteins - immunology; Viral Envelope Proteins - isolation & purification
• ISSN: 0889-2229
• DOI: 10.1089/aid.1988.4.51

We partially purified the human immunodeficiency virus (HIV) glycoprotein gp41 from infected H9 cells by immunoaffinity chromatography using a column containing the M25 monoclonal antibody (diMarzo-Veronese et al., 1985). A pH 11.5 buffer worked best for eluting the glycoprotein from this column. The eluted gp41 was used in a sensitive slot blot immunoassay to detect antibodies to HIV in human sera and to prepare rabbit polyclonal antibodies and the 41-1S mouse monoclonal antibody. These antibodies reacted with gp41 in immunoprecipitation and in Western blot assays, but did not neutralize HIV in a syncytium-forming microassay. A pH 2.5 buffer was found to be the most effective solution for eluting gp41 from a 41-1S monoclonal antibody column.