Epitope mapping of rat neutralizing monoclonal antibody against human immunodeficiency virus type-1 by a phage peptide library: comparison with ELISA using synthetic peptides.
We generated a rat monoclonal antibody (mAb W#10) with the ability to neutralize human immunodeficiency virus type 1IIIB (HIV-1IIIB) infection. The epitope recognized by mAb W#10 was defined as R-I-Q-R-G-P-G by enzyme-linked immunosorbent assay (ELISA) with the use of synthetic peptides. The filamentous phage clones displaying random 15-amino-acid peptides on the amino terminus of the pIII coat protein reacting with mAb W#10 were identified with affinity and immunological selection procedures. Thirteen out of 16 selected phage clones contained the G-X-G-R-X-F sequence in the coat protein region representing significant homology to a part of conserved G-P-G-R-A-F sequence in the V3 loop of various HIV-1 strains. In addition, the phage clones included the G-X-G sequence in the sequence detected by synthetic peptides as the recognition site. The selected phage clones were stained by mAb W#10 specifically and were able to compete with mAb binding to cells expressing viral antigens.
Ichiyama
K
K
Department of Microbiology, Tokyo Medical and Dental University School of Medicine, Japan.
Ishikawa
D
D
Tanaka
Y
Y
Kashiwa
T
T
Koyanagi
Y
Y
Handa
S
S
Yamashita
A
A
Fukushi
M
M
Yamamoto
N
N
Taki
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001666:Binding Sites, Antibody; D002478:Cells, Cultured; D003090:Coliphages / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018604:Epitope Mapping / Q000379:methods*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007668:Kidney; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019151:Peptide Library; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis* / Q000276:immunology*; D051381:Rats
D000911:Antibodies, Monoclonal; D019151:Peptide Library; D010455:Peptides