Table Of ContentMONOCLONAL
ANTIBODIES
Hybridomas: A New Dimension
in Biological Analyses
MONOCLONAL
ANTIBODIES
Hybridomas: A New Dimension
in Biological Analyses
Edited by
Roger H. Kennett
and
1LhonnasJ~cKearn
University of Pennsylvania School of Medicine
Philadelphia, Pennsylvania
and
Kathleen B. Bechtol
The Wistar Institute
Philadelphia, Pennsylvania
Plenum Press . New York and London
Library of Congress Cataloging in Publication Data
Main entry under title:
Monoclonal antibodies.
Includes index.
1. hnmunoglobulins. 2. Gammopathies, Monoclonal. 3. Cell hybridization.
L Kennett, Roger H. IL McKearn, Thomas J. m. Bechtoi,!Kathleen B. [DNLM:
1. Antibodies. 2. Clone cells - hnmunology. QW575 M751]
QRI86.7.M66 599.02'93 80-15118
ISBN-13: 978-1-4615-7507-8 e-ISBN-13: 978-1-4615-7505-4
DOl: 10.1007/978-1-4615-7505-4
©1980 Plenum Press, New York
Softcover reprint of the hardcover 1s t edition 1980
A Division of Plenum Publishing Corporation
227 West 17th Street, New York, N.Y. 10011
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without written permission from the Publisher
In Memoriam
J.
WILLIAM MELLMAN
Scientist, Pediatrician, and Friend
May 7, 1928-February 27, 1980
Contributors
C. ABRAMS, Department of Human Genetics, University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
KATHLEEN B. BECHTOL, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania
19104
YEHUDIT BERGMAN, Howard Hughes Medical Institute Laboratories and Department of Medicine,
Stanford University Medical Center, Stanford, California 94305
IRWIN D. BERNSTEIN, Pediatric Oncology Program of the Fred Hutchinson Cancer Research
Center, Seattle, Washington 98104, and Department of Pediatrics, University of Washington,
Seattle, Washington 98195
PAUL H. BLACK, Department of Medicine, Harvard Medical School, and Massachusetts General
Hospital, Boston, Massachusetts 02114
J.
ROGER BRIDEAU, MRC Cellular Immunology Unit, Sir William Durtn School of Pathology,
University of Oxford, Oxford OXI 3RE, England
SHERRI BROWN, Howard Hughes Medical Institute Laboratories and Department of Medicine,
Stanford University Medical Center, Stanford, California 94305
W. NEAL BURNETTE, Tumor Virology Program of the Fred Hutchinson Cancer Research Center,
Seattle, Washington 98104
M.P. CANCRO, Department of Pathology, University of Pennsylvania, School of Medicine, Philadel
phia, Pennsylvania 19104
EDWARD A. CLARK, Department of Genetics, University of Washington, Seattle, Washington 98195
BRIAN CLEVINGER, Department of Microbiology and Immunology, Washington University School
of Medicine, St. Louis, Missouri 63110
M. C. COSEO, Department of Human Genetics, University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
JOSEPH DAVIE, Department of Microbiology and Immunology, Washington University School of
Medicine, St. Louis, Missouri 63110
KATHLEEN DENIS, Department of Human Genetics, University of Pennsylvania, School of Medi
cine, Philadelphia, Pennsylvania 19104
BETTY DIAMOND, Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New
York 10461
vii
viii CONTRIBUTORS
JEANETTE DILLEY, Howard Hughes Medical Institute Laboratories and Department of Medicine,
Stanford University Medical Center, Stanford, California 94305
EDGAR G. ENGLEMAN, Department of Pathology, Stanford University Medical Center, Stanford,
California 94305
FRANK W. FITCH, The Committee on Immunology and the Department of Pathology, University of
Chicago, Chicago, Illinois 60637
MARK E. FRANKEL, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
WALTER GERHARD, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
JAMES W. GoDING, Department of Genetics, Stanford University School of Medicine, Stanford,
California 94305
RICHARD A. GoLDSBY, Department of Chemistry, University of Maryland, College Park, Maryland
20742
ROGERS GRIFFITH, Department of Microbiology and Immunology, Washington University School
of Medicine, St. Louis, Missouri 63110
J. B. HAAS, Department of Human Genetics, University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
EDGAR HABER, Department of Medicine, Harvard Medical School, and Massachusetts General
Hospital, Boston, Massachusetts 02114
DANIEL HANSBURG, Department of Microbiology and Immunology, Washington University School
of Medicine, St. Louis, Missouri 63110
H. HARRIS, Department of Human Genetics, University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
HANS HENGARTNER, Basel Institute for Immunology, CH-4005 Basel5,Switzerland
LEONARD A. HERZENBERG, Department of Genetics, Stanford University School of Medicine,
Stanford, California 94305
LEROY HOOD, Division of Biology, California Institute of Technology, Pasadena, California 91125
L.L. HOUSTON, Tumor Virology Program or'the Fred Hutchinson Cancer Research Center, Seat
tle, Washington 98104
J. C. HOWARD, Department of Pathology, University of Pennsylvania, School of Medicine, Philadel
phia, Pennsylvania 19104
ZDENKA L. JONAK, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
E. A. JONES, Genetics Laboratory, Department of Biochemistry, University of Oxford, Oxford OXI
3QU, England. Present address: Division of Immunology, Stanford University Medical School,
Stanford, California 94305
ROGER H. KENNETT, Department of Human Genetics, University of Pennsylvania, School of
Medicine, Philadelphia, Pennsylvania 19104
DENNIS M. KUNMAN, Department of Pathology, University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
NORMAN KUNMAN, Department of Cellular and Developmental Immunology, Scripps Clinic and
Research Foundation, La Jolla, California 92037
HILARY KOPROWSKI, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
CONTRIBUTORS IX
LoIS A. LAMPSON, Department of Anatomy, University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
JEFFREY A. LEDBETTER, Department of Genetics, Stanford University School of Medicine, Stan
ford, California 94305
RONALD LEVY, Howard Hughes Medical Institute Laboratories and Department of Medicine,
Stanford University Medical Center, Stanford, California 94305
A. DwIGHT LoPES, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
MARK E. LOSTROM, Tumor Virology Program of the Fred Hutchinson Cancer Research Center,
Seattle, Washington 98104
J.
THOMAS McKEARN, Department of Pathology, Divisions of Research Immunology and Labora
tory Medicine, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania
19104
BRIAN McMASTER, Pediatric Oncology Program of the Fred Hutchinson Cancer Research Center,
Seattle, Washington 98104
W. ROBERT McMASTER, MRC Cellular Immunology Unit, Sir William Dunn School of Pathology,
University of Oxford, Oxford OXI 3RE, England
DAVID H. MARGULIES, Department of Cell Biology, Albert Einstein College of Medicine, Bronx,
New York 10461
DoNALD W. MASON, MRC Cellular Immunology Unit, Sir William Dunn School of Pathology,
University of Oxford, Oxford OXI 3RE, England
TOMASO MEO, Basel Institute for Immunology, CH-4005 Basel 5, Switzerland
CHRISTINE MOLINARO, Department of Cellular and Developmental Immunology, Scripps Clinic
and Research Foundation, La Jolla, California 92037
EDITH MULLER, Basel Institute for Immunology, CH-4005 Basel 5, Switzerland
ROBERT C. NOWINSKI, Tumor Virology Program of the Fred Hutchinson Cancer Research Center,
Seattle, Washington 98104, and Department of Microbiology, University of Washington, Seattle,
Washington 98195
PAUL V. O'DONNELL, Memorial Sloan-Kettering Cancer Center, New York, New York 10021
BARBARA A. OSBORNE, Department of Genetics, Stanford University Medical Center, Stanford,
California 94305
RICHARD A. POLIN, Department of Pediatrics, University of Pennsylvania, School of Medicine, and
The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104
MATTHEW D. ScHARFF, Department of Cell Biology, Albert Einstein College of Medicine, Bronx,
New York 10461
JAMES SCHILLING, Division of Biology, California Institute of Technology, Pasadena, California
91125
LINDA SHERMAN, Department of Cellular and Developmental Immunology, Scripps Clinic and
Research Foundation, La Jolla, California 92037
C. A. SLAUGHTER, Department of Human Genetics, University of Pennsylvania, School of Medi
cine, Philadelphia, Pennsylvania 19104
DAWN E. SMILEK, Department of Pathology, Division of Research Immunology, University of
Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
x CONTRIBUTORS
E. SOLOMON, Genetics Laboratory, Department of Biochemistry, University of Oxford, Oxford
OXI 3QU, England. Present address: Imperial Cancer Research Fund, P.O. Box 123, Lincoln's
Inn Fields, London WC2A 3PX, England
TIMOTHY A. SPRINGER, Pathology Department, Harvard Medical School, Boston, Massachusetts
02115
LOUIS STAUDT, Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
MARY R. STONE, Tumor Virology Program of the Fred Hutchinson Cancer Research Center,
Seattle, Washington 98104
MILTON R. TAM, Tumor Virology Program of the Fred Hutchinson Cancer Research Center,
Seattle, Washington 98104
TAKESHI TOKUHISA, Department of Genetics, Stanford University School of Medicine, Stanford,
California 94305
MICHAEL WEBB, MRC Cellular Immunology Unit, Sir William Dunn School of Pathology, Univer
sity of Oxford, Oxford OXI 3RE, England
ROBERT A. H. WHITE, MRC Cellular Immunology Unit, Sir William Dunn School of Pathology,
University of Oxford, Oxford OXI 3RE, England
TADEUSZ WIKTOR, Wi star Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
ALAN F. WILLIAMS, MRC Cellular Immunology Unit, Sir William Dunn School of Pathology,
University of Oxford, Oxford OX 1 3RE, England
DALE E. YELTON, Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New
York 10461
JONATHAN YEWDELL, Wi star Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104
VINCENT R. ZURAWSKI, JR., Department of Medicine, Harvard Medical School, and Massachusetts
General Hospital, Boston, Massachusetts 02114
Preface
On August 7, 1975, Kohler and Milstein published in Nature (256:495) a report
describing "Continuous cultures of fused cells secreting antibody of predefined
specificity." Their report has become a classic and has already had a profound
effect on basic and applied research in biology and medicine. By the time the first
Workshop on Lymphocyte Hybridomas (Current Topics in Microbiology and Im
munology 81, 1978) was held on April 3-5, 1978, in Bethesda, Maryland, investi
gators from many laboratories had made hybrids between plasmacytomas and
spleen cells from immunized animals and had obtained monoclonal antibodies
reacting with a broad variety of antigenic determinants.
At the time Kohler and Milstein introduced this new technology, the editors
of this volume were involved in the production of antisera against differentiation
antigens (K.B.B.), histocompatibility antigens (T.]. McK.), and human tumor
associated antigens (R.H.K.). Because of the potential usefulness of monoclonal
antibodies in these areas, we each began production of hybridomas and analysis
of the resulting monoclonal reagents. One of the most interesting aspects of
participation in the early stages of the development and application of hybrid
oma technology has been observing how the implications of the initial observa
tions gradually spread first among the practitioners of immunology and immu
nogenetics, and then to other areas of the biological sciences, such as
developmental biology, biochemistry, human genetics, and cell and tumor
biology.
Besides providing a novel way of obtaining homogeneous antibodies that
will be useful in many areas of biology, Kohler and Milstein's achievement also
provides a prototype for the use of hybrid cells to "trap" clones of cells perform
ing a specialized function as transformed cell lines producing a specific product
or performing a specific function in vitro. When one considers that the produc
tion of specific immunoglobulins may be only the first of several such functions
that could be "immortalized" by cell fusion, then it appears likely that the full
implications of Kohler and Milstein's report may still be far from full realization.
This volume is designed to present an overview of the production and uses
xi
Xli PREFACE
of monoclonal antibodies and to facilitate their application to other areas of
biology. It is intended for advanced undergraduates, graduate students, and
scientists in the biological and medical sciences. We do not presume to consider
the volume comprehensive, but have attempted to include examples of many of
the approaches and applications that have been developed to exploit this
technology .
Roger H. Kennett
J.
Thomas McKearn
Kathleen B. Bechtol
Philadelphia
Description:On August 7, 1975, Kohler and Milstein published in Nature (256:495) a report describing "Continuous cultures of fused cells secreting antibody of predefined specificity. " Their report has become a classic and has already had a profound effect on basic and applied research in biology and medicine.
