[Epitope mapping of HIV-1 using phage-display random peptide library and the purified IgG from HIV patient].
OBJECTIVE
By using phage display random peptide library, the B cell epitope of HIV protein was studied.


METHODS
The library displaying random dodecamers was biopanned first with human total IgG antibodies against HIV-1, and then non-specific phages were subtracted by purified IgG from non-HIV sera. After three rounds of screening, the positive phages were tested by ELISA for their reactivity with HIV(+)-IgG and HIV(-)-IgG antibodies. Phage that showed positive reactivity with HIV(+)-IgG, but negative to HIV(-)-IgG, were selected and their displayed peptides were determined by DNA sequencing.


RESULTS
All the 13 positive clones sequenced displayed five kinds of peptides (SPKCLGKLLCAF, THQCLGKLQCGV, SCSAKFTCTTQI, KSDCSARFMCSV, DCLKQWACEWSR) that have homology to the HIV-I gp4l (602GCSGKLICTIN613).


CONCLUSIONS
This method demonstrated there is a dominant epitope in the region of HIV-1 gp4l and can be used in the research of the B cell epitope of HIV protein.
Du
Y
Y
Department of Applied Molecular Biology, Institute of Microbiology and Epidemiology, Academy of Military Medicine, Beijing 100071, China.
Hou
L
L
Li
J
J
D001435:Bacteriophages; D018604:Epitope Mapping / Q000379:methods*; D018985:Epitopes, B-Lymphocyte / Q000032:analysis*; D015488:HIV Antigens / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology* / Q000302:isolation & purification; D019151:Peptide Library
D018985:Epitopes, B-Lymphocyte; D015488:HIV Antigens; D007074:Immunoglobulin G; D019151:Peptide Library