Antibody Phage Display: Methods and Protocols

241 Pages·2009·2.44 MB·English

by  Beatrix Kotlan

M M B ™
ETHODS IN OLECULAR IOLOGY
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Antibody Phage Display
Methods and Protocols
Second Edition
Edited by
Robert Aitken
Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, UK
Editor
Dr. Robert Aitken
Institute of Biomedical and Life Sciences
University of Glasgow
Glasgow, UK
[email protected]
ISSN 1064-3745 e-ISSN 1940-6029
ISBN 978-1-60327-301-5 e-ISBN 978-1-60327-302-2
DOI 10.1007/978-1-60327-302-2
Springer Dordrecht Heidelberg London New York
Library of Congress Control Number: 2009926045
© Humana Press, a part of Springer Science+Business Media, LLC 2009
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Preface
When the first edition of Antibody Phage Display: Methods and Protocols was published,
I and my co-editor Philippa O’Brien expressed the hope that the book would form a
resource of real value for scientists who had followed the development of phage display
technology through its first decade. The large number of citations to the book’s content
and the success of its sales provided some assurance that we met that goal.
The aim of providing a resource that will guide readers in the design and execution
of experiments based around antibody phage display holds true for this second edition;
it goes alongside the hope that the methods will inspire investigators with the possibilities
presented by this and related technologies. These include the huge range of methods for
isolating recombinant antibodies from phage display libraries, how the targets recognised
by antibodies of interest can be identified, the identification and exploitation of antibodies
that can enter cells and bind to cytosolic targets, and novel approaches to the expression
of recombinant antibodies.
This is a field of biology in which innovation is the norm: in assembling contributions,
I have therefore tried to stimulate the reader’s imagination as much as guide their practice
in the laboratory.
I am indebted to all the contributing authors for sharing their expertise with the wider
scientific community. I also pay thanks to my graduate students past and present with
whom I have enjoyed exploring an area of biology that is forever moving forward.
Glasgow, UK Robert Aitken
v
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Antibody Phage Display: Overview of a Powerful Technology
that Has Quickly Translated to the Clinic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Beatrix Kotlan and Mark C. Glassy
2 Design and Construction of Synthetic Phage-Displayed Fab Libraries. . . . . . . . . . 17
Jenny Bostrom and Germaine Fuh
3 Affinity Isolation of Antigen-Specific Circulating B Cells for
Generation of Phage Display-Derived Human Monoclonal Antibodies . . . . . . . . . 37
Henrik J. Ditzel
4 Construction of Phage Antibody Repertoires
from the Blood of West Nile Virus-Infected Donors . . . . . . . . . . . . . . . . . . . . . . . 45
Mark Throsby and John de Kruif
5 Anti-b GP-I and Anti-prothrombin Antibodies Generated
2
by Phage Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Javier Cabiedes and Antonio R. Cabral
6 The Generation and Selection of Single-Domain,
V Region Libraries from Nurse Sharks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Martin F. Flajnik and Helen Dooley
7 The Isolation of scFvs Against Small Target Molecules . . . . . . . . . . . . . . . . . . . . . 83
Iain Shaw and Marian Kane
8 Display and Selection of scFv Antibodies on HEK-293T Cells . . . . . . . . . . . . . . . 99
Mitchell Ho and Ira Pastan
9 Isolation of scFvs that Inhibit the NS3 Protease of Hepatitis C Virus
by a Combination of Phage Display and a Bacterial Genetic Screen. . . . . . . . . . . . 115
Limor Nahary, Alla Trahtenherts, and Itai Benhar
10 Guided Selection Methods Through Chain Shuffling . . . . . . . . . . . . . . . . . . . . . . 133
Bao Guo-qiang and He Xian-Li
11 Sequential Antigen Panning for Selection of Broadly Cross-Reactive
HIV-1-Neutralizing Human Monoclonal Antibodies . . . . . . . . . . . . . . . . . . . . . . 143
Mei-Yun Zhang and Dimiter S. Dimitrov
12 Selection of Antibodies Able to Rapidly Enter Mammalian Cells. . . . . . . . . . . . . . 155
Marie-Alix Poul
13 Identification of the Specificity of Isolated Phage Display
Single-Chain Antibodies Using Yeast Two-Hybrid Screens . . . . . . . . . . . . . . . . . . 165
Nicolaj Rasmussen and Henrik J. Ditzel
14 Generation of Bispecific and Tandem Diabodies. . . . . . . . . . . . . . . . . . . . . . . . . . 177
Sergey M. Kipriyanov
vii
viii Contents
15 Intrabody Expression in Eukaryotic Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
Laurence Guglielmi and Pierre Martineau
16 High-Level Periplasmic Expression and Purification of scFvs. . . . . . . . . . . . . . . . . 205
Sergey M. Kipriyanov
17 Expression of Single-Chain Fv Fragments in E. coli Cytoplasm . . . . . . . . . . . . . . . 215
Laurence Guglielmi and Pierre Martineau
18 High-Level Expression of a Phage Display-Derived scFv in Pichia pastoris. . . . . . . 225
Leonardo M. Damasceno, Frank Lee, Gerd Ritter, Lloyd Old,
and Carl Batt
Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237
Contributors
CARL BATT (cid:129) Department of Food Science, Cornell University, Ithaca, NY, USA
ITAI BENHAR (cid:129) George S. Wise Faculty of Life Sciences, Tel-Aviv University,
Ramat Aviv, Israel
JENNY BOSTROM (cid:129) Genentech Inc., South San Francisco, CA, USA
ANTONIO R. CABRAL (cid:129) Instituto Nacional de Ciencias Médicas y Nutrición Salvador
Zubirán, Mexico City, Mexico
JAVIER CABIEDES (cid:129) Instituto Nacional de Ciencias Médicas y Nutrición Salvador
Zubirán, Mexico City, Mexico
LEONARDO M. DAMASCENO (cid:129) Sanofi Pasteur, Swiftwater, PA, USA
DIMITER S. DIMITROV (cid:129) National Cancer Institute at Frederick, National Institutes of
Health, Frederick, MD, USA
HENRIK J. DITZEL (cid:129) Institute of Medical Biology, University of Southern Denmark,
Odense, Denmark
HELEN DOOLEY (cid:129) University of Maryland School of Medicine, Baltimore, MD, USA
MARTIN F. FLAJNIK (cid:129) University of Maryland School of Medicine, Baltimore, MD, USA
GERMAINE FUH (cid:129) Genentech Inc., South San Francisco, CA, USA
MARK C. GLASSY (cid:129) Integrated Medical Sciences Association Foundation, San Diego,
CA, USA
LAURENCE GUGLIELMI (cid:129) Université Montpellier I, Montpellier F-34298, France
BAO GUO-QIANG (cid:129) Tangdu Hospital, Xi’an 710038, Shaanxi Province, China
MITCHELL HO (cid:129) National Cancer Institute, National Institutes of Health, Bethesda,
MD, USA
MARIAN KANE (cid:129) National Centre for Biomedical Engineering Science, National
University of Ireland, Galway, Ireland
SERGEY M. KIPRIYANOV (cid:129) Affi tech AS, N-0349 Oslo, Norway
BEATRIX KOTLAN (cid:129) National Institute of Oncology, Budapest, Hungary
JOHN DE KRUIF (cid:129) Merus Biopharmaceuticals, Utrecht, The Netherlands
FRANK LEE (cid:129) Ludwig Institute for Cancer Research, New York, NY, USA
PIERRE MARTINEAU (cid:129) Université Montpellier I, Montpellier F-34298, France
LIMOR NAHARY (cid:129) George S. Wise Faculty of Life Sciences, Tel-Aviv University, Ramat
Aviv, Israel
LLOYD OLD (cid:129) Ludwig Institute for Cancer Research, New York, NY, USA
IRA PASTAN (cid:129) National Cancer Institute, National Institutes of Health, Bethesda,
MD, USA
MARIE-ALIX POUL (cid:129) UMR CNRS 8113, Ecole Normale Supérieure de Cachan, Cachan,
France
NICOLAJ RASMUSSEN (cid:129) Institute of Medical Biology, University of Southern Denmark,
Odense, Denmark
GERD RITTER (cid:129) Ludwig Institute for Cancer Research, New York, NY, USA
ix
x Contributors
IAIN SHAW (cid:129) National Centre for Biomedical Engineering Science, National University
of Ireland, Galway, Ireland
MARK THROSBY (cid:129) Merus Biopharmaceuticals, Utrecht, The Netherlands
ALLA TRAHTENHERTS (cid:129) George S. Wise Faculty of Life Sciences, Tel-Aviv University,
Ramat Aviv, Israel
HE XIAN-LI (cid:129) Tangdu Hospital, Xi’an 710038, Shaanxi Province, China
MEI-YUN ZHANG (cid:129) National Cancer Institute at Frederick, National Institutes of
Health, Frederick, MD, USA
Chapter 1
Antibody Phage Display: Overview of a Powerful
Technology that Has Quickly Translated to the Clinic
Beatrix Kotlan and Mark C. Glassy
Summary
Antibody-based immunologic reagents are useful for identifying, isolating, or eliminating cells with
particular characteristics related to different diseases. Phage display is a highly valuable technique for
antibody selection related to this purpose. In brief, a diverse group of antibody genes prepared from a
patient or generated in vitro are inserted into a phagemid vector or the phage genome so that when the
protein is expressed, it becomes anchored on the surface of the phage by fusion to a coat protein. A
diverse library of recombinant antibodies is generated in this way and can then be exposed or panned on
the antigen of interest, typically, this being a molecule associated with a particular pathological condition.
Phage that carry proteins or peptides bind preferentially to the target and can thus be isolated from the
library. The viruses that are recovered in this way also carry the gene for the binding moiety facilitating its
over-expression or manipulation. Recent reviews highlight key milestones in the development of antibody
libraries and their screening by phage display, and the impact of these technologies on drug discovery
seems assured.
Key words: Antibody fragment , Phage display library , Target antigen , Specific binding
1. Introduction
Since its introduction almost 20 years ago ( 1– 4 ) , phage display
technology has completely transformed approaches to the analysis
of biomedical problems. Initial drawbacks and skepticism were
quickly overcome by growing evidence of the technology’s strong
and well-defined scientific process. The ability to rapidly select and
analyze binding interfaces has been its most powerful application.
Robert Aitken (ed.), Antibody Phage Display, Second Edition, Methods in Molecular Biology, vol. 562
DOI 10.1007/978-1-60327-302-2_1, © Humana Press, a part of Springer Science + Business Media, LLC 2009
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