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In order to develop a reliable and inexpensive serodiagnostic method, gag protein of human immunodeficiency virus type 1 (HIV-1), p24 (HIV-gag; bp 730-1419)was cloned into an expression vector pCT10 with a sequence encoding a hydroxylamine cleavage site and with a part of lacZ gene (lacZ": 834 bp) as a fusion partner. Overexpression of lacZ"-p24 was induced in E. coli and the p24 fusion protein was purified to homogeneity by centrifugation, hydroxylamine cleavage, and two steps of column chromatography (ion-exchange chromatography and gel filtration chromatography). Western blot analysis and enzyme-linked immunosorbant assay (ELISA) using the purified p24 peptide as an antigen showed high sensitivity and specificity for detecting HIV-1 antibodies in testing human sera when used with a previously purified gp41 epitope which we purified previously. The mixed antigens showed higher sensitivities and specificities on ELISA test than with the gp41 epitope alone [Sohn, M. J., Cho, S. H., Jang, W. H., Chong, Y. H., Nham, S. U., and Lee, Y. I. (1993) J. Virol. Methods 41, 93-100]. These results suggest that simple and rapid purification of recombinant core antigen will provide a valuable resource of HIV-1 serodiagnostics for screening large groups of blood donors to prevent HIV-1 infection.
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