Selection and identification of single domain antibody fragments from camel heavy-chain antibodies

被引:630
作者
Ghahroudi, MA [1 ]
Desmyter, A [1 ]
Wyns, L [1 ]
Hamers, R [1 ]
Muyldermans, S [1 ]
机构
[1] FREE UNIV BRUSSELS VIB, B-1640 RHODE ST GENESE, BELGIUM
关键词
antibody; phage display; single domain antibody fragment; VH; camel;
D O I
10.1016/S0014-5793(97)01062-4
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Functional heavy-chain gamma-immunoglobulins lacking light chains occur naturally in Camelidae, We now show the feasibility of immunising a dromedary cloning the repertoire of the variable domains of its heavy-chain antibodies and panning, leading to the successful identification of minimum sized antigen binders, The recombinant binders are expressed web in E. coli, extremely stable, highly soluble, and react specifically and with high affinity to the antigens, This approach can be viewed as a general route to obtain small binders with favourable characteristics and valuable perspectives as modular building blocks to manufacture multispecific or multifunctional chimaeric proteins. (C) 1997 Federation of European Biochemical Societies.
引用
收藏
页码:521 / 526
页数:6
相关论文
共 31 条
[1]   ASSEMBLY OF COMBINATORIAL ANTIBODY LIBRARIES ON PHAGE SURFACES - THE GENE-III SITE [J].
BARBAS, CF ;
KANG, AS ;
LERNER, RA ;
BENKOVIC, SJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (18) :7978-7982
[2]  
BIRD RE, 1989, SCIENCE, V241, P423
[3]   Interactions of protein antigens with antibodies [J].
Davies, DR ;
Cohen, GH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (01) :7-12
[4]   Single antibody domains as small recognition units: Design and in vitro antigen selection of camelized, human VH domains with improved protein stability [J].
Davies, J ;
Riechmann, L .
PROTEIN ENGINEERING, 1996, 9 (06) :531-537
[5]   CAMELISING HUMAN-ANTIBODY FRAGMENTS - NMR-STUDIES ON VH DOMAINS [J].
DAVIES, J ;
RIECHMANN, L .
FEBS LETTERS, 1994, 339 (03) :285-290
[6]   ANTIBODY VH DOMAINS AS SMALL RECOGNITION UNITS [J].
DAVIES, J ;
RIECHMANN, L .
BIO-TECHNOLOGY, 1995, 13 (05) :475-479
[7]   An antibody VH domain with a lox-Cre site integrated into its coding region: Bacterial recombination within a single polypeptide chain [J].
Davies, J ;
Riechmann, L .
FEBS LETTERS, 1995, 377 (01) :92-96
[8]   Crystal structure of a camel single-domain V-H antibody fragment in complex with lysozyme [J].
Desmyter, A ;
Transue, TR ;
Ghahroudi, MA ;
Thi, MHD ;
Poortmans, F ;
Hamers, R ;
Muyldermans, S ;
Wyns, L .
NATURE STRUCTURAL BIOLOGY, 1996, 3 (09) :803-811
[9]   A CONVENIENT ENZYME-LINKED IMMUNOSORBENT-ASSAY FOR TESTING WHETHER MONOCLONAL-ANTIBODIES RECOGNIZE THE SAME ANTIGENIC SITE - APPLICATION TO HYBRIDOMAS SPECIFIC FOR THE BETA-2-SUBUNIT OF ESCHERICHIA-COLI TRYPTOPHAN SYNTHASE [J].
FRIGUET, B ;
DJAVADIOHANIANCE, L ;
PAGES, J ;
BUSSARD, A ;
GOLDBERG, M .
JOURNAL OF IMMUNOLOGICAL METHODS, 1983, 60 (03) :351-358
[10]   MEASUREMENTS OF THE TRUE AFFINITY CONSTANT IN SOLUTION OF ANTIGEN-ANTIBODY COMPLEXES BY ENZYME-LINKED IMMUNOSORBENT-ASSAY [J].
FRIGUET, B ;
CHAFFOTTE, AF ;
DJAVADIOHANIANCE, L ;
GOLDBERG, ME .
JOURNAL OF IMMUNOLOGICAL METHODS, 1985, 77 (02) :305-319