title: Epitope mapping of rat neutralizing monoclonal antibody against human immunodeficiency virus type-1 by a phage peptide library: comparison with ELISA using synthetic peptides.
abstract: 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.
authors.affiliation: Department of Microbiology, Tokyo Medical and Dental University School of Medicine, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ichiyama
authors.forename: D
authors.initials: D
authors.lastname: Ishikawa
authors.forename: Y
authors.initials: Y
authors.lastname: Tanaka
authors.forename: T
authors.initials: T
authors.lastname: Kashiwa
authors.forename: Y
authors.initials: Y
authors.lastname: Koyanagi
authors.forename: S
authors.initials: S
authors.lastname: Handa
authors.forename: A
authors.initials: A
authors.lastname: Yamashita
authors.forename: M
authors.initials: M
authors.lastname: Fukushi
authors.forename: N
authors.initials: N
authors.lastname: Yamamoto
authors.forename: T
authors.initials: T
authors.lastname: Taki
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal; D019151:Peptide Library; D010455:Peptides