       Document 0103
 DOCN  M9640103
 TI    Epitope mapping of anti-HIV and anti-HCV monoclonal antibodies and
       characterization of epitope mimics using a filamentous phage peptide
       library.
 DT    9604
 AU    Grihalde ND; Chen YC; Golden A; Gubbins E; Mandecki W; Aging and
       Degenerative Disease Department, Abbott Laboratories,; North Chicago, IL
       60064, USA.
 SO    Gene. 1995 Dec 12;166(2):187-95. Unique Identifier : AIDSLINE
       GENBANK/L39948
 AB    A large filamentous phage library (1 x 10(9) clones) displaying random
       30-amino-acid (aa) sequences on the N terminus of the pIII coat protein
       was constructed and characterized. Clones in the library were affinity
       selected for binding to monoclonal antibodies (mAb) against two viral
       antigens, the HIV gp120 protein and the HCV core protein. The obtained
       aa sequences precisely identified the epitopes recognized by the mAb.
       Binding of peptide-carrying phages to the Ab was demonstrated by ELISA,
       Western blot and the surface plasmon resonance (SPR) method. The
       mAb-specific peptides were transferred via genetic techniques onto the N
       terminus of Escherichia coli alkaline phosphatase (AP). When fused to
       the enzyme, the peptides maintained their ability to bind their
       respective mAb, indicating that the peptides contained the necessary
       contact residues for binding. The affinity of the peptides was estimated
       to be 100 nM by SPR. A comparison is presented of the relative
       affinities of phage-derived peptides to the native viral epitopes also
       displayed on the AP scaffold. The approach of transferring epitopes from
       phage to AP for further evaluation should be applicable to many other
       mAb or receptors.
 DE    Acid Phosphatase/GENETICS  Amino Acid Sequence  Antibodies,
       Monoclonal/IMMUNOLOGY  Antibody Affinity  Antibody Specificity  Base
       Sequence  DNA Primers/CHEMISTRY  Epitope Mapping  Gene Library  Genetic
       Vectors  Hepatitis B Core Antigens/*IMMUNOLOGY  Hepatitis C
       Viruses/IMMUNOLOGY  HIV Antigens/*IMMUNOLOGY  HIV Envelope Protein
       gp120/*IMMUNOLOGY  HIV-1/IMMUNOLOGY  Molecular Sequence Data
       Recombinant Fusion Proteins/IMMUNOLOGY  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

