Monoclonal hybridoma antibodies : techniques and applications

240 Pages·2018·13.89 MB·English

by  Hurrell

Monoclonal Hybridoma
Antibodies:
Techniques
and
Applications
Editor
John G. R. Hurrell, Ph.D.
Head, Immunochemistry R & D Group
Commonwealth Serum Laboratories
Parkville, Victoria
Australia
CRC Press
(cpp)
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Library of Congress Cataloging in Publication Data
Main entry under title:
Monoclonal hybridoma antibodies.
Bibliography: p.
Includes index.
1. Antibodies, Monoclonal. I. Hurrell,
John G. R., 1949-
QR186.85.M663 616.07’93 81-18129
ISBN 0-8493-6511-2 AACR2
A Library of Congress record exists under LC control number: 81018129
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PREFACE
The landmark paper of Kohler and Milstein (Nature (London), 256, 495, 1975) de­
scribing the production and characterization of the first somatic cell hybrids capable of
indefinite production of antibody of predetermined specificity, heralded a new era in
biology and initiated a virtual “avalanche” of scientific literature. The potential of the
somatic cell hybridization (hybridoma) technique was quickly realized by investigators
in many areas of biology and applied to a wide variety of antigens. The technique has
been refined and many alternative protocols developed, but all with the same basic core
element, the fusion of primed splenocytes with biosynthetically defective myeloma
cells.
The first section of this volume was aimed to provide a comprehensive review of the
many varied and often empirically derived techniques and procedures currently in use
to produce monoclonal hybridoma cell lines and to characterize the antibodies secreted.
This goal has been achieved with the chapter contributed by Zola and Brooks who, as
each step in the process of hybridoma production and antibody characterization is re­
viewed, have provided an experimental procedure found to be satisfactory in their lab­
oratory. Investigators about to begin a hybridoma program are thus presented with back­
ground information for the rapid assessment of alternative procedures. The advantages,
faults, and known pitfalls of each method are discussed and, if confused by the nu­
merous procedures in the literature, the investigator may choose the experimental pro­
cedure of the authors to serve as a starting point.
The second section of this volume was designed to provide a review of areas in which
monoclonal hybridoma antibodies have been of particular advantage. This is a rapidly
advancing field which could not be thoroughly reviewed in a single volume. The par­
ticular examples reviewed were selected from the broad areas of soluble antigens (so-
matotrophic hormones, cardiac myosin, alpha fetoprotein, and carcinoembryonic an­
tigen), viral antigens (influenza virus and herpes simplex virus), parasite antigens,
tumor associated human tissue antigens, and nontumor tissue antigens (macrophage
membrane, lymphocyte membrane, and histocompatibility antigens). These specific
applications of monoclonal hybridoma antibodies are intended as general reviews as
well as illustrating and expanding on many of the points brought out by Zola and Brooks
in the first chapter.
It is apparent that the surface has only just been broken regarding the information
that monoclonal hybridoma antibodies will provide. The future will see the widespread
development and use of monoclonal hybridoma antibodies for routine immunotherapy
and diagnosis, and for furthering our understanding of the molecular biology of the cell
and of disease processes. There is no doubt that this is an exciting time to be a biological
scientist!
JOHN G. R. HURRELL
Parkville, Australia
December 1981
THE EDITOR
John G. R. Hurrell, Ph.D., is Head of Immunochemistry Research and Development
at the Commonwealth Serum Laboratories in Parkville, Australia. He is also an As­
sociate in the Department of Pathology and Immunology, Monash University, Mel­
bourne.
Dr. Hurrell graduated from the University of Melbourne in 1971 with a B.Sc. in
Chemistry and Biochemistry. In 1974, after 3 years as a Research and Development
scientist at the Commonwealth Serum Laboratories, he returned to the University of
Melbourne to study under Professor Sidney J. Leach in the Department of Biochem­
istry. Dr. Hurrell received a Ph.D. in 1977. After a postdoctoral year at the Common­
wealth Serum Laboratories, Dr. Hurrell was awarded a Lulbright Lellowship in the
Cellular and Molecular Research Laboratory, Massachusetts General Hospital, Boston.
Lollowing a short period as a Visiting Scientist at Centocor Inc., Philadelphia, Dr.
Hurrell returned to Australia in July 1980 to take up his present position.
CONTRIBUTORS
Rosemary Betts, B.Sc. J. Ivanyi, Ph.D.
Research Associate Head, Department of Experimental
Department of Pathology Immunobiology
University of Melbourne The Wellcome Research Laboratories
Parkville, Victoria Beckenham, Kent
Australia England
D. A. Brooks, B.Sc. Hugo A. Katus, M.D.
Ph.D. Student Fellow in Internal Medicine
Department Clinical Immunology School of Medicine
Flinders Medical Centre University of Heidelberg
Bedford Park, South Australia Heidelberg, West Germany
Paul Ehrlich, Ph.D.
Ban-An Khaw, Ph.D.
Research Associate
Assistant Professor
Department of Medicine
Department of Pathology
College of Physicians & Surgeons
Harvard Medical School
Columbia University
Associate Biochemist
New York, New York
Cellular and Molecular Research Lab
Massachusetts General Hospital
George M. Georgiou, B.Sc. Hons Boston, Massachusetts
Research Fellow
Royal Children’s Hospital Research
Foundation Hilary Koprowski, M.D.
Royal Children’s Hospital Director
Parkville, Victoria The Wistar Institute
Australia Philadelphia, Pennsylvania
Edgar Haber, M.D. Herbert Z. Kupchik, Ph.D.
Professor of Medicine Assistant Professor
Harvard Medical School Department of Microbiology
Chief, Cardiac Unit Boston University Medical Center
Massachusetts General Hospital Member, Hubert H. Humphrey
Boston, Massachusetts Cancer Research Center
Boston University
C. S. Hosking, M.D. Member, Special Scientific Staff
Director, Department of Immunology Mallory Institute of Pathology
Royal Children’s Hospital Boston, Massachusetts
Parkville, Victoria
Australia
W. G. Laver, Ph.D.
Senior Fellow
John G. R. Hurrell, Ph.D. Microbiology Department
Head, Immunochemistry R & D John Curtin School of Medical
Group Research
Commonwealth Serum Laboratories Australian National University
Parkville, Victoria Canberra City
Australia Australia
Gary R. Matsueda, Ph.D. Lenore Pereira, Ph.D.
Assistant Professor of Pathology Research Specialist
Harvard Medical School California State Department of Public
Assistant in Biochemistry Health Services
Massachusetts General Hospital Viral and Rickettsial Disease
Boston, Massachusetts Laboratory
Berkeley, California
Ian F. C. McKenzie, M.D., Ph.D.
Timothy A. Springer, Ph.D.
Department of Pathology
Chief, Membrane Immunochemistry
University of Melbourne
Sidney Farber Cancer Institute
Parkville, Victoria
Assistant Professor of Pathology
Australia
Harvard Medical School
Boston, Massachusetts
Graham F. Mitchell, Ph.D.
Laboratory Head Zenon Steplewski, M.D.,
Laboratory of Immunoparasitology Ph.D., D.Sc.
The Walter and Eliza Hall Institute The Wistar Institute
of Medical Research Philadelphia, Pennsylvania
Melbourne, Victoria
Australia
H. Zola, Ph.D.
Chief Hospital Scientist
Kenneth F. Mitchell, Ph.D., Flinders Medical Centre
F.R.S.A. Senior Lecturer, Immunology
Assistant Professor Flinders University of South
The Wistar Institute Australia
Philadelphia, Pennsylvania Bedford Park, South Australia
Vincent R. Zurawski, Jr., Ph.D.
Vice President and Technical
Director
Centocor
Philadelphia, Pennsylvania
TABLE OF CONTENTS
Chapter 1
Techniques for the Production and Characterization of Monoclonal
Hybridoma Antibodies ......................................................................................................... 1
H. Zola and D. Brooks
Chapter 2
Analysis of Monoclonal Antibodies to Human Growth Hormone and
Related Proteins .................................................................................................................... 59
J. Ivanyi
Chapter 3
Antibodies to Alphafetoprotein and Carcinoembryonic Antigen Produced
by Somatic Cell Fusion ...................................................................................................... 81
Herbert Z. Kupchik
Chapter 4
Monoclonal Antibodies Specific for Cardiac Myosin: In Vivo and In Vitro
Diagnostic Tools in Myocardial Infarction ..................................................................... 91
Edgar Haber, Hugo A. Katus, John G. Hurrell, Gary R. Matsueda,
Paul Ehrlich, Vincent Zurawski, Jr., and Ban-An Khaw
Chapter 5
The Use of Monoclonal Antibodies to Investigate Antigenic Drift in
Influenza Virus ........................................................................................................................103
W. G. Laver
Chapter 6
Monoclonal Antibodies to Herpes Simplex Viruses 1 and 2 .........................................119
Lenore Pereira
Chapter 7
Hybridomas in Immunoparasitology ..................................................................................139
Graham F. Mitchell
Chapter 8
Hybridoma Antibodies Specific for Human Tumor Antigens .......................................151
Kenneth F. Mitchell, Zenon Steplewski, and Hilary Koprowski
Chapter 9
Murine Macrophage Differentiation Antigens Defined by Monoclona Antibodies 169
Timothy A. Springer
Chapter 10
Application of Monoclonal Antibodies to the Study of Human Lymphocyte
Surface Antigens ......................................................................................................................177
C. S. Hosking and G. M. Georgiou
Chapter 11
Monoclonal Antibodies to the Major Histocompatability Antigens............................193
Rosemary L. Betts and Ian F. C. McKenzie
Index .........................................................................................................................................223
1
Chapter 1
TECHNIQUES FOR THE PRODUCTION AND CHARACTERIZATION OF
MONOCLONAL HYBRIDOMA ANTIBODIES
H. Zola and D. Brooks
TABLE OF CONTENTS
I. Introduction ................................................................................................................... 4
II. Prerequisites to Starting a Hybridoma Project....................................................... 5
A. Major Equipment ........................................................................................... 5
1. Cabinet for Sterile Work .................................................................. 5
2. Gassed Incubator ............................................................................... 7
3. Liquid Nitrogen Storage Facility .................................................... 7
B. Minor Equipment ............................................................................................ 7
C. Expertise ............................................................................................................. 8
III. Materials and Media .................................................................................................... 8
A. Materials/Chemicals to be Ordered ............................................................ 8
1. Cell Culture Medium: RPMI 1640, or Alternative
(Dulbeccos; Iscoves).......................................................................... 8
2. Fetal Calf Serum: Selected Batch .................................................. 8
3. HAT Selective Medium Components ............................................ 8
4. Polyethylene Glycol (PEG) ............................................................. 8
5. Dimethyl Sulfoxide ........................................................................ 9
B. Working Solutions ........................................................................................... 9
1 Medium ................................................................................................. 9
2. HAT Medium ...................................................................................... 9
3. HT Medium ........................................................................................ 9
4. Phosphate-Buffered Saline (PBS) .................................................. 9
5. Gey’s Hemolytic Medium .................................................................10
6. PEG/DMSO Fusing Agent Contains 42% PEG W/V
and 15% DMSO (V/V) .....................................................................10
7. Alternative Antibiotics or Antifungal Agents .........................,. 11
IV. Antigens and Immunization Schedules .....................................................................11
A. Introduction ........................................................................................................11
B. Effectiveness of Immunization .......................................................................11
C. Enrichment of Antibody-Producing Cells.....................................................11
D. Cell Membrane Antigens .................................................................................12
E. Improving the Proportion of Hybrids Specific for the
“Desired” Antigen .........................................................................................12
F. Soluble Antigens ...............................................................................................14
G. Mouse Strain .....................................................................................................14