USE OF A GENE-TARGETED PHAGE DISPLAY RANDOM EPITOPE LIBRARY TO MAP AN ANTIGENIC DETERMINANT ON THE BLUETONGUE VIRUS OUTER CAPSID PROTEIN VP5

被引:69
作者
WANG, LF [1 ]
DUPLESSIS, DH [1 ]
WHITE, JR [1 ]
HYATT, AD [1 ]
EATON, BT [1 ]
机构
[1] ONDERSTEPOORT VET INST, ONDERSTEPOORT, SOUTH AFRICA
关键词
BLUETONGUE VIRUS; EPITOPE MAPPING; LIBRARY; PHAGE SURFACE DISPLAY; DNASE I DIGESTION; RANDOM EXPRESSION;
D O I
10.1016/0022-1759(94)00235-O
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We describe the use of a gene-targeted random epitope library for the mapping of antigenic determinants. A DNA clone encoding the target antigen was digested randomly with DNase I to generate a population of DNA fragments of different sizes and sequences. After size fractionation, small DNA fragments (100-200 bp) were isolated and cloned into the phage expression vector fUSE2 to form an expression library displaying random polypeptide sequences as fusion proteins at the N terminus of the phage gene III protein. This library, termed a gene-targeted random epitope library to distinguish it from totally random synthetic epitope libraries, was then screened by affinity selection for recombinant phages which were specifically bound by the antibody of interest. Using this approach, we have mapped a monoclonal antibody (mAb)-defined epitope on the bluetongue virus outer capsid protein VP5. This epitope is not accessible on the intact virus surface, but is recognised by the immune system of sheep and cattle during virus infection. Although the example given here utilised a DNA fragment of known sequence and the library was screened for a mAb-defined epitope, the strategy described should be equally applicable to genes of unknown sequence and for screening of epitopes using polyclonal antibodies. The approach can also be extended to identify immunodominant epitope from much more complex genome-targeted random epitope library for virus, bacteria and eukaryotic organisms. Other applications of recombinant phages expressing defined immunodominant epitopes include serodiagnosis and vaccine development.
引用
收藏
页码:1 / 12
页数:12
相关论文
共 51 条
[1]   ANTIGENIC STRUCTURES OF PROTEINS - THEIR DETERMINATION HAS REVEALED IMPORTANT ASPECTS OF IMMUNE RECOGNITION AND GENERATED STRATEGIES FOR SYNTHETIC MIMICKING OF PROTEIN-BINDING SITES [J].
ATASSI, MZ .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1984, 145 (01) :1-20
[2]   IDENTIFICATION OF A HEXAPEPTIDE THAT MIMICS A CONFORMATION-DEPENDENT BINDING-SITE OF ACETYLCHOLINE-RECEPTOR BY USE OF A PHAGE-EPITOPE LIBRARY [J].
BALASS, M ;
HELDMAN, Y ;
CABILLY, S ;
GIVOL, D ;
KATCHALSKIKATZIR, E ;
FUCHS, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (22) :10638-10642
[3]   THE ANTIGENIC STRUCTURE OF PROTEINS - A REAPPRAISAL [J].
BENJAMIN, DC ;
BERZOFSKY, JA ;
EAST, IJ ;
GURD, FRN ;
HANNUM, C ;
LEACH, SJ ;
MARGOLIASH, E ;
MICHAEL, JG ;
MILLER, A ;
PRAGER, EM ;
REICHLIN, M ;
SERCARZ, EE ;
SMITHGILL, SJ ;
TODD, PE ;
WILSON, AC .
ANNUAL REVIEW OF IMMUNOLOGY, 1984, 2 :67-101
[4]   ENGINEERED IRON OXIDE-ADHESION MUTANTS OF THE ESCHERICHIA-COLI PHAGE-LAMBDA RECEPTOR [J].
BROWN, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (18) :8651-8655
[5]   EXPRESSION POLYMERASE CHAIN-REACTION FOR THE INVITRO SYNTHESIS AND EPITOPE MAPPING OF AUTOANTIGEN - APPLICATION TO THE HUMAN THYROTROPIN RECEPTOR [J].
BURCH, HB ;
NAGY, EV ;
KAIN, KC ;
LANAR, DE ;
CARR, FE ;
WARTOFSKY, L ;
BURMAN, KD .
JOURNAL OF IMMUNOLOGICAL METHODS, 1993, 158 (01) :123-130
[6]   ASSOCIATION OF BLUETONGUE VIRUS GENE SEGMENT-5 WITH NEUROINVASIVENESS [J].
CARR, MA ;
DEMATTOS, CC ;
DEMATTOS, CA ;
OSBURN, BI .
JOURNAL OF VIROLOGY, 1994, 68 (02) :1255-1257
[7]   EPITOPE DISCOVERY USING PEPTIDE LIBRARIES DISPLAYED ON PHAGE [J].
CORTESE, R ;
FELICI, F ;
GALFRE, G ;
LUZZAGO, A ;
MONACI, P ;
NICOSIA, A .
TRENDS IN BIOTECHNOLOGY, 1994, 12 (07) :262-267
[8]   SCREENING FOR RECEPTOR LIGANDS USING LARGE LIBRARIES OF PEPTIDES LINKED TO THE C-TERMINUS OF THE LAC REPRESSOR [J].
CULL, MG ;
MILLER, JF ;
SCHATZ, PJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (05) :1865-1869
[9]   PEPTIDES ON PHAGE - A VAST LIBRARY OF PEPTIDES FOR IDENTIFYING LIGANDS [J].
CWIRLA, SE ;
PETERS, EA ;
BARRETT, RW ;
DOWER, WJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (16) :6378-6382
[10]   ANTIBODY-ANTIGEN COMPLEXES [J].
DAVIES, DR ;
PADLAN, EA ;
SHERIFF, S .
ANNUAL REVIEW OF BIOCHEMISTRY, 1990, 59 :439-473