Table Of ContentAdvisory Board:
ELMER L. B E C K ER
UNIVERSITY OF CONNECTICUT HEALTH CENTER
FRANK J. DIXON
SCRIPPS CLINIC AND RESEARCH FOUNDATION
HERMAN EISEN
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
PIERRE GRABAR
THE PASTEUR INSTITUTE, PARIS
JOHN H. HUMPHREY
ROYAL POST-GRADUATE MEDICAL SCHOOL
HAMMERSMITH HOSPITAL, LONDON
ELVIN A. KABAT
COLUMBIA-PRESBYTERIAN MEDICAL CENTER
HENRY G. K U N K EL
THE ROCKEFELLER UNIVERSITY
MACLYN MCCARTY
THE ROCKEFELLER UNIVERSITY
JACQUES OUDIN
THE PASTEUR INSTITUTE, PARIS
DAVID PRESSMAN
ROSWELL PARK MEMORIAL INSTITUTE
Methods in
IMMUNOLOGY
and IMMUNOCHEMISTRY
Edited by
CURTIS A. W I L L I A MS
THE STATE UNIVERSITY OF NEW YORK
PURCHASE, NEW YORK
MERRILL W. CHASE
THE ROCKEFELLER UNIVERSITY
NEW YORK, NEW YORK
Volume IV
Agglutination, Complement, Neutralization, and Inhibition
1977
ACADEMIC PRESS New York Son Francisco London
A SUBSIDIARY OF HARCOURT BRACE JOVANOVICH, PUBLISHERS
COPYRIGHT © 1977, BY ACADEMIC PRESS, INC.
ALL RIGHTS RESERVED.
NO PART OF THIS PUBLICATION MAY BE REPRODUCED OR
TRANSMITTED IN ANY FORM OR BY ANY MEANS, ELECTRONIC
OR MECHANICAL, INCLUDING PHOTOCOPY, RECORDING, OR ANY
INFORMATION STORAGE AND RETRIEVAL SYSTEM, WITHOUT
PERMISSION IN WRITING FROM THE PUBLISHER.
ACADEMIC PRESS, INC.
Ill Fifth Avenue, New York, New York 10003
United Kingdom Edition published by
ACADEMIC PRESS, INC. (LONDON) LTD.
24/28 Oval Road, London NWl
library of Congress Cataloging in Publication Data
Williams, Curtis Alvin, Date
Methods in immunology and immunochemistry.
Includes bibliographies.
CONTENTS.-v. 1. Preparation of antigens and
antibodies.-v. 2. Physical and chemical methods,
-v. 3. Reactions of antibodies with soluble
antigens.-v. 4. Agglutination, complement,
neutralization, and inhibition.
1. Immunology. 2. Immunochemistry-Technique.
I. Chase, Merrill W., joint author. II. Title.
QR181.W68 615'.37 67-22779
ISBN 0-12-754404-6
PRINTED IN THE UNITED STATES OF AMERICA
Contributors to Volume IV
Numbers in parentheses indicate the pages on which the authors^ contributions begin.
E. R. ARQUILLA (41), Department of Pathology, College of Medicine,
University of California, Irvine, California
RICHARD C. BLINKOFF (91)/^ The Rockefeller University, New York,
New York
Ç. E. BOND (226),*'' National Cancer Institute, Bethesda, Maryland
WILLIAM C. BOYD (19),f Boston University School of Medicine, Bos
ton, Massachusetts
J. BOZICEVICH (121),t Biomedics Research Laboratories, Inc., Divi
sion of Litton Industries, Kensington, Maryland
MERRILL W. CHASE (54, 61, 127, 136, 166, 180, 194,198, 200, 226, 238,
241, 242, 250, 253, 254, 256, 258, 261, 266, 267, 269, 271, 275,
282, 285, 293, 296, 303, 407), The Rockefeller University, New York,
New York
B. CINADER (313), Institute of Immunology, Departments of Medical
Genetics, Medical Biophysics, and Clinical Biochemistry, Medical
Sciences Building, University of Toronto, Ontario, Canada
HARVEY R. COLTEN (226), Department of Pediatrics, Harvard
Medical School, Boston, Massachusetts
R. R. A. COOMBS (49, 54, 56), Department of Pathology, University of
Cambridge, Cambridge, England
MARY B. GIBBS (1, 2, 4, 73), Department of Immunology, Walter
Reed Army Institute of Research, Washington, D.C.
0. GÖTZE (219), Department of Molecular Immunology, Scripps
Clinic and Research Foundation, La Jolla, California
* Present address: 830 Broadway, New York, New York.
**Present address: Electro-Nucleonics Laboratories, Inc., Bethesda, Maryland.
t Present address: 1241 Prospect Street, La Jolla, California.
t Present address: 6810 Hillmead Road, Bethesda, Maryland.
xi
Xll CONTRIBUTORS TO VOLUME IV
JOSEPH HAIMOVICH (386), Department of Chemical Immunology,
The Weizmann Institute of Science, Rehovot, Israel
PETER M. HENSON (271), Department of Experimental Pathology,
Scripps Clinic and Research Foundation, La Jolla, California
LOUIS G. HOFFMANN (127, 137,167, 173, 225, 227, 228, 232), Depart
ment of Microbiology, College of Medicine, State University of
Iowa, Iowa City, Iowa
ROLLIN D. HOTCHKISS (88), The Rockefeller University, New York,
New York
M. A. JESAITIS (375), The Rockefeller University, New York, New
York
MAURICE LANDY (26),* National Institute of Allergy and Infectious
Diseases, Bethesda, Maryland
IRWIN H. LEPOW (180, 183, 186, 201, 242), Department of Medicine,
University of Connecticut Health Center, Farmington, Connecticut
PHILIP LEVINE (9, 12), Immunohematology Division, Ortho Research
Foundation, Raritan, New Jersey
STEPHEN D. LITWIN (115),t Department of the Army Headquarters,
U.S. Army Medical Research Laboratory, Fort Knox, Kentucky
MANFRED M. MAYER (127, 137, 167, 173, 225, 227, 228, 229, 232,
235, 238, 241, 256, 261, 267), Department of Microbiology, The
Johns Hopkins School of Medicine, Baltimore, Maryland
P. L. MOLLISON (49, 54), Wright-Fleming Institute of Microbiology,
St. Mary's Hospital Medical School, London, England
HANS J. MÜLLER-EBERHARD (127, 176, 180, 190, 205, 209, 212, 217,
219, 220, 223, 250, 260, 263, 265, 269), Department of Molecular
Immunology, Scripps Clinic and Research Foundation, La Jolla,
California
ERWIN NETER (76, 89) ,ß Departments of Pediatrics and Microbio
logy, State University of New York at Buffalo, Buffalo, New York
* Present Address: Schweizerisches Forschungs Institut, 7270 Davos, Switzerland,
t Present address: Division of Human Genetics, Cornell University Medical College,
New York, New York.
$ Present address: Department of Bacteriology, Children's Hospital, Buffalo, New
York.
CONTRIBUTORS ÔĎ VOLUME IV XÜl
JACK PENSKY (242),» Department of Medicine, Case Western Re
serve University, Cleveland, Ohio
M. RAYNAUD (277, 283, 288, 289, 292, 294, 301, 302, 310) ,t Institut
Pasteur, Annexe de Garches, Garches, France
E. H. RELYVELD (277, 283, 288, 289, 292, 294, 301, 302, 310), Institut
Pasteur, Annexe de Garches, Garches, France
MICHAEL SELA (386), Department of Chemical Immunology, The
Weizmann Institute of Science, Rehovot, Israel
SIDNEY SHULMAN (94), Sperm Antibody Laboratory, Department
of Microbiology, Departments of Urology and of Obstetrics and
Gynecology, New York Medical College, New York, New York
GEORG F. SPRINGER (14, 67), Department of Microbiology, North
western University, Evanston Hospital, Evanston, Illinois
ABRAM B. STAVITSKY (30, 47), Department of Microbiology, Case
Western Reserve University School of Medicine, Cleveland, Ohio
MARGARET TREACY (9,12), Immunohematology Division, Ortho Re
search Foundation, Raritan, New Jersey
CURTIS A. WILLIAMS (127,136, 166, 180, 194, 198, 200, 226, 238, 241,
242, 250, 253, 254, 256, 258, 261, 266, 267, 269, 271),t The Rockefeller
University, New York, New York
N. D. ZINDER (375), The Rockefeller University, New York, New York
•Present address: Cleveland Veterans' Hospital, 10701 East Boulevard, Cleveland,
Ohio 44106.
t Deceased.
t Present address: The State University of New York, Purchase, New York.
Preface
In planning an open-ended treatise on methodology in immunochemistry
and immunology, we considered the needs not only of the specialist but
of those whose primary concern in other areas of biology may suggest
application of immunological tools. A rigid format was adopted for the
first five volumes, a choice which has caused unexpected problems in
publication. Volumes published to date have been very well received.
Volume I is concerned with typical preparative methods employed in
handling antigens, antibodies, and laboratory animals. Volume II presents
general chemical and physiochemical methods used in immunological re
search. Volume III deals with antigen-antibody and hapten-antibody
reactions in vitro, in free solution, and in gels. Volume IV covers direct
and indirect agglutination reactions, complement-fixation procedures,
hemolytic intermediates, isolation of complement components, comple
ment-related proteins, antisera to complement components, and neutral
ization reactions (toxins, enzymes, bacteria, bacteriophages). Volume V
deals with antigen-antibody reactions in vivo : anaphylaxis, Arthus
reactions, tolerance, immune suppression with chemical agents, radiation
effects, phagocytosis and clearance, antibody synthesis in vitro, immuno-
histological methods, and applied electron microscopy.
Our aim has been to open our colleagues' notebooks to bring together
detailed procedures that are hard to retrieve from the original literature
and to provide indexes which can be used with confidence. Contributors
were asked to include not only the details of procedures they had found
most satisfactory in their own laboratory, but also critical remarks on
common pitfalls and interpretation of results, references to alternative
methods, and mention of applications to other problems. While not all
topics are easily suited to this format, we feel that insofar as our general
objectives are achieved, these volumes represent high potential energy.
Some important general topics as well as many specific methods are to
be covered in future volumes including hypersensitivity, immunity to
infection, transplantation, and immunogenetics.
We again express our appreciation to the advisory editors.
CURTIS A. WILLIAMS
MERRILL W. CHASE
XV
Contents of Other Volumes
VOLUME I
1. ANTIGENS
Proteins
H. F. Deutsch
Bacterial Antigens
M. J. How, H. Koffler, R. M. Krause, Ę. C. Milner, S. I. Morse, E. Ribi, A. M.
Staub, L. W. Wannamaker
Blood Group Antigens
J. KoScielak, W. T. J. Morgan
Viruses
M. A. Jesaitis, E. H. Lennette, R. MacLeod, N. J. Schmidt, N. D. Zinder
Conjugated and Synthetic Antigens
S. M. Reiser, F. Borek, F. Edel, Ç. Í. Eisen, Â. F. Erlanger, S. Fuchs, S. Lieber-
man, V. Likhite, J. R. Little, R. Marks, A. Nisonoff, C. W. Parker, 0. J. Plescia,
A. Sehon, M. Sela, S. W. Tanenbaum
Preparation and Testing of Lipids for Immunological Study
L. Graf, M. M. Rapport
2. PRODUCTION OF ANTISERUM
A. A. Benedict, M. W. Chase, E. A. Kabat
3. PURIFICATION OF ANTIBODY
Preparation of Immunoglobulin
J. R. Cann, H. F. Deutsch, J. L. Fahey, H. H. Fudenberg
Preparation of Specific Antibody
D. H. Campbell, H. N. Eisen, W. Gray, E. A. Kabat, N. R. Klinman,
S. Kochwa, J. R. Little, R. E. Rosenfield, E. S. Simms, S. J. Singer, B. T. Tozer,
N. Weliky
XViii CONTENTS OF OTHER VOLUMES
4. LABELING OF ANTIGENS AND ANTIBODIES
H. N. Claman, J. S. Garvey, K. C. Hsu, D. Pressman, J. Radovich, 0. A. Roholt,
D. W. Talmage, C. A. Williams
5. METHODS USED IN STUDIES OF THE STRUCTURE OF
IMMUNOGLOBULINS
G. M. Edelman, J. J. Marchalonis
VOLUME II
0. ELECTROPHORESIS
J. R. Cann, B. J. Davis, J. Kohn, H. J. Müller-Eberhard, L. Ornstein, C. K.
Osterland, J. Porath, M. D. Poulik, S. Raymond, J. Uriel, C. A. Wilhams
7. ULTRACENTRIFUGATION (Preparative and Analytical)
K. M. Cowan, R. Trautman
8. DIALYSIS AND ULTRAFILTRATION
L. C. Craig
9. PREPARATIVE CHROMATOGRAPHY
S. Hjerten, T. P. King
10. OPTICAL ANALYSIS
H. Donahue, C. A. Leone, J. R. Little
11. RADIOISOTOPE ANALYSIS
J. S. Garvey, R. E. Ritts, Jr.
12. CHEMICAL ANALYSES
M. W. Chase, L. Gidez, L. Levine, B. Mach, W. T. Murakami, C. A. Wilhams
APPENDIX I. SPECIAL PROPERTIES OF PROTEINS AND SMALL MOLECULES OF
IMMUNOLOGICAL INTEREST
H. Donahue, J. R. Little
APPENDIX II. BUFFERS
M. W. Chase
APPENDIX III. POWER BLOCK ELECTROPHORESIS
C. K. Osterland
CONTENTS OF OTHER VOLUMES XIX
VOLUME III
13. PRECIPITATION REACTIONS
John J. Cebra, Richard S. Farr, Charles A. Leone, 1. W. Li, Paul H. Maurer,
Abraham G. Osier, Curtis A. Williams
14. PRECIPITATION ANALYSIS BY DIFFUSION IN GELS
Frederick Aladjem, A. C. Allison, Elmer L. Becker, Merrill W. Chase,
Minchin Clarke, Alfred J. Crowle, D. A. Darcy, G. C. Easty, I. Finger,
Joseph F. Heremans, Stellan Hjerten, G. M. Hochwald, J. John, J. Kohn,
Felix Milgrom, J. Munoz, Jacques Oudin, Alfred Poison, M. D. Poulik,
John R. Freer, Jr., B. Russell, Harry Smith, G. J. Thorbecke, Jose Uriel,
Curtis A. Williams
15. HAPTEN REACTIONS AND KINETICS OF INTERACTION WITH ANTIBODIES
John J. Cebra, Richard A. Dammkoehler, Walter B. Dandliker, Herman W.
Eisen, A. Froese, Tom L. Gallagher, Fred Karush, Sally S. Karush, James E.
McGuigan, A. H. Sehon
APPENDIX I. IDENTIFICATION OF MULTIPLE ANTIBODY COMPONENTS BY
RADIOIMMUNOELECTROPHORESIS AND RADIOIMMUNODIFFUSION
Yasuo Yagi
VOLUME V
19. ANAPHYLAXIS
K. F. Austen, M. Bach, W. E. Brockelhurst, Ě. W. Chase, M. J. Conrad,
E. J. Coulson, G. A. Feigen, Ĺ. Haber, L. L. Layton, R. P. Orange, A. G. Osier,
Z. Ovary, P. A. Shore, R. P. Siraganian, S. H. Snyder, S. Spector, J. Spragg,
D. J. Stechschulte, R C. Talamo, K. M. Taylor, M. D. Valentine, M. Walzer,
Â. υ. Wintroub
20. THE ARTHUS AND RELATED REACTIONS
C. G. Cochrane
21. ANALYSIS OF ANTIGEN-ANTIBODY REACTIONS IN CORNEA
F. G. Germuth, Jr.
22. IMMUNOLOGICAL TOLERANCE
W. 0. Weigle
