Partial purification of native HIV transmembrane protein gp41: generation of polyclonal and monoclonal antibodies.
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.
Zweig
M
M
Program Resources, Inc., NCI-Frederick Cancer Research Facility, MD.
Showalter
S D
SD
Bladen
S V
SV
Gilden
R V
RV
Arthur
L O
LO
Robey
W G
WG
Nara
P L
PL
Fischinger
P J
PJ
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D002846:Chromatography, Affinity; D006678:HIV / Q000032:analysis* / Q000276:immunology; D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41; D006801:Humans; D009500:Neutralization Tests; D012191:Retroviridae Proteins / Q000276:immunology / Q000302:isolation & purification*; D014759:Viral Envelope Proteins / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins