HIV-1 envelope protein gp120 expression by secretion in E. coli: assessment of CD4 binding and use in epitope mapping.
A non-glycosylated form of the HIV-1 envelope protein gp120 and four truncated derivatives have been expressed as non-fused secreted products in the periplasmic space of E. coli. We show that the full length molecule, whilst folded and soluble, fails to bind to CD4 consistent with other work that suggests an essential role for carbohydrate in gp120 function. In addition, when used in conjunction with the truncated derivatives, rapid epitope mapping of anti-gp120 monoclonal antibodies is achieved using both Western-blot and ELISA formats.
Morikawa
Y
Y
NERC Institute of Virology and Environmental Microbiology, Oxford, U.K.
Moore
J P
JP
Jones
I M
IM
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology*; D015704:CD4 Antigens / Q000276:immunology*; D003001:Cloning, Molecular; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics*; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015704:CD4 Antigens; D000939:Epitopes; D015699:HIV Envelope Protein gp120