Making and using antibodies : a practical handbook

446 Pages·2013·18.23 MB·English

by  Howard

SECOND EDITION
MAK ING
AND USING
ANTIBODIES
A Practical Handbook
Edited by
Gary C. Howard
Matthew R. Kaser
SECOND EDITION
MAKING
AND USING
ANTIBODIES
A Practical Handbook
Edited by
Gary C. Howard
Matthew R. Kaser
Boca Raton London New York
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Contents
Preface......................................................................................................................vii
Acknowledgment ......................................................................................................ix
Editors .......................................................................................................................xi
Contributors ...........................................................................................................xiii
Chapter 1 Antibodies ............................................................................................1
Matthew R. Kaser and Gary C. Howard
Chapter 2 Antigens and Adjuvants .......................................................................7
Paul Algate, Jory Baldridge, and Sally P. Mossman
Chapter 3 Production of Polyclonal Antibodies ..................................................33
Lon V. Kendall
Chapter 4 Purification and Characterization of Antibodies ...............................67
Joseph P. Chandler
Chapter 5 Production of Monoclonal Antibodies ...............................................97
Kathleen C. F. Sheehan
Chapter 6 Quantitative Production of Monoclonal Antibodies ........................119
David A. Fox and Elizabeth M. Smith
Chapter 7 Making Antibodies in Bacteria ........................................................151
Frederic A. Fellouse and Sachdev S. Sidhu
Chapter 8 In Vitro Selection of Aptamers .........................................................173
Tianjiao Wang, Xiangyu Cong, and Marit Nilsen-Hamilton
Chapter 9 Chemical and Proteolytic Modification of Antibodies ....................207
George P. Smith
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vi Contents
Chapter 10 Western Blots and Other Applications .............................................275
Lee Bendickson and Marit Nilsen-Hamilton
Chapter 11 Immunohistochemical Methods .......................................................303
José A. Ramos-Vara
Chapter 12 Immunoelectron Microscopy ...........................................................343
Sara E. Miller and David N. Howell
Chapter 13 Flow Cytometry ...............................................................................371
Steven B. McClellan
Chapter 14 ELISAs .............................................................................................385
John Chen and Gary C. Howard
Chapter 15 Humanization of Antibodies ............................................................395
Juan Carlos Almagro, Sreekumar Kodangattil, and Jian Li
Chapter 16 Antibodies in the Future: Challenges and Opportunities .................421
Gary C. Howard and Matthew R. Kaser
Index ......................................................................................................................429
Preface
Antibodies are some of the most extraordinary proteins ever to evolve during the
history of life on Earth. They protect us from a wide range of infectious diseases
and cancers. For science, these “magic bullets” have become an indispensable tool.
In biology, they have been a key component of the surge in fundamental knowledge
that has occurred in the last quarter century. They are relatively easy to make and
use, have great flexibility in applications, and are cost-effective for most labs. As
well as being generated for use in conventional immune response research, they have
been used in many other different aspects of basic research, including identification
of modified proteins and enzymes, of transcription factors, of membrane proteins, as
well as immunoprecipitation of unknown proteins having a known epitope. In medi-
cine, multiple vaccines have led to the control (at least in the developed world) of
many infectious diseases, including polio, mumps, measles, and chicken pox, as well
as the eradication of smallpox, and are being increasingly used to combat cancer.
We hope this methods book will be useful to biomedical researchers and students,
although new methods for making and using antibodies will certainly be found.
In this edition, we added chapters on the use of aptamers and the humanization of
antibodies. Nevertheless, the many current applications—ELISAs, western blotting,
immunohistochemistry, and flow cytometry—are so powerful that they will remain
critical to biomedical science for a considerable period.
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vii

Acknowledgment
We thank the contributors to this volume. Their professional knowledge, excellent
writing, and enthusiastic support made this possible outcome of this book.
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