Detection of inhibition of HIV-1 protease activity by an enzyme-linked immunosorbent assay (ELISA).
An ELISA is described for the detection of HIV-1 protease activity using an immobilized gag-related polyprotein as substrate. Proteolytic activity was demonstrated with either bacterial lysates expressing HIV-1 protease or purified protease. No cleavage was observed with a protein preparation from control bacteria not expressing HIV-1 protease. Under these conditions the aspartyl-type protease inhibitor, pepstatin A, was found to inhibit HIV-1 protease cleavage by > 90% at a concentration of 0.1 mM. This assay may be a useful tool for the study of both synthetic and natural inhibitors of HIV-1 protease.
Department of Internal Medicine, Medical Academy of Magdeburg, Germany.
H W
HW
Mansfeld
S
S
Schulz
G
G
Grütz
R
R
von Baehr
S
S
Ansorge
D004797:Enzyme-Linked Immunosorbent Assay; D016655:HIV Core Protein p24 / Q000378:metabolism; D015700:HIV Envelope Protein gp41 / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000032:analysis*; D015497:HIV-1 / Q000201:enzymology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41; D017320:HIV Protease Inhibitors; D011993:Recombinant Fusion Proteins