Monoclonal Antibody Technology: The Production and Characterization of Rodent and Human Hybridomas PDF
Preview Monoclonal Antibody Technology: The Production and Characterization of Rodent and Human Hybridomas
Monoclonal an tibody technology LABORATORY TECHNIQUES IN BIOCHEMISTRY AND MOLECULAR BIOLOGY Volume 13 Edited by R.H. BURDON - Department of Biochemistry, University of Glusgo w P.H. van KNIPPENB,ERG - Department of Biochemistry, University of Leiden Advisory board P. BORST - University of Amsterdam D.C. BURKE - University of Warwick P.B. GARLAND - University of Dundee M. KATES - University of Ottawa W. SZYBALSKI - University of Wisconsin H.G. WlTTMAN - Max-Planck Institut fur Molekulare Genetik, Berlin ELSE V I E R AMSTERDAM . NEW YORK . OXFORD MONOCLONAL ANTIBODY TECHNOLOGY THE PRODUCTION AND CHARACTERIZATION OF RODENT AND HUMAN HYBRIDOMAS Ailsa M. Campbell Department of Biochemistry, University of Glasgow, Glasgow GI2 8QQ (Great Britain) 1984 ELSEVIER AMSTERDAM . NEW YORK . OXFORD 1984, Elsevier Science Publishers B. V. AN rights reserved. No part of thispublication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, mechanical, photocopying, recording or otherwise, without the priorpermission of the copyright owner. However, this book has been registered with the Copyright Clearance Center, Inc. Consent is given for copying pages for personal of internal use of specific clients, This consent is given on the condition that the copier pay through the Center the perpage fee stated below for copying beyond that permitted by the U.S.C opyright Law. The appropriate fee should be forwarded with a copy of front and back of the title page of the book to the Copyright Clearance Center, Salem, MA 0197). This consent does not extend to other kinds of copying, such as for general distribution, resale, advertising and promotional purposes, or for creating new works. Special written permission must be obtained from the publisher for such copying. The per page fee code for this book is 0-444-80575-3 (pocket edition): 84/$0.80; 0-444-80592-3 (library edition): 84/$0.80. series: 0-7204-4200-1: 84/$0.80. ISBN 0-444-80575-3 (pocket edition) 0-444-80592-3 (library edition) series: 0-7204-4200-1 Published by: ELSEVIER SCIENCE PUBLISHERS PO BOX 211 1000 AE AMSTERDAM THE NETHERLANDS Sole distributors for the USA and Canada: ELSEVIER SCIENCE PUBLISHING COMPANY, INC. 52 VANDERBILT AVENUE NEW YORK, NY 10017 USA Library of Congress Cataloging in Publication Data Campbell, Ailsa M. Monoclonal antibody technology. Includes index. 1. Antibodies, Monoclonal. 2. Hybridomas. 3. Biotechnology. I. Title. CDNIM: 1. Antibodies, Monoclonal. 2. Hybidomas. 3. Immunologic Technics. w1 LA232K v.13 / Qw 575 C187mI ~86.85c.3 6 1984 615 * .37 84-10322 ISBN 0-444-80592-3 ISBN 0-444-80575-3 (pbk.) This book is the pocket edition of Volume 13, of the series ‘Laboratory Techniques in Biochemistry and Molecular Biology’ Printed in The Netherlands Preface This book is essentially a laboratory manual based on the protocols and general advice currently available in our own department. It was written, for the two groups of people who have consulted us extensive- ly over the past few years. The first were competent biochemists who were interested in the potential of monoclonal antibodies as reagents either to study basic biochemistry or to develop better clinical assay systems. The second were clinicians with an interest in the potential of monoclonal antibodies for the detection and potential therapy of various clinical conditions who had sufficient technical assistance to feel that they might embark on monoclonal antibody production themselves using material from their own patients. The main questions raised by discussions with these two groups were related to the possibility of obtaining the monoclonal antibody (or panel of monoclonal antibodies) required in a period of 1 - 2 years or indeed at all. Many of the general multi-author review articles written by those who had been successful give the impression that monoclonal antibodies may be produced against any epitope of any molecule, however impure and the practical problems that such an approach might encounter had to be analysed. An example might be, how sensible is it to try and raise a panel of human monoclonal antibodies to a tumour of limited allogeneic cross reaction, of which samples are available at a level of lo7 tumour cells every six months. Another might be the practicability of raising rodent monoclonal antibodies to a molecule which is only available in a highly impure state. A third might be the likelihood of making rodent monoclonal antibodies of a chosen specificity and affinity to a small peptide vi MONOCLONAL ANTIBODY TECHNOLOGY hormone. Many of the projects suggested seemed unlikely to succeed for technical reasons while some had considerable potential. The book attempts to guide the reader through what may be possible and what is less likely to succeed in the limited time period available to most laboratories. Those projects which had the most potential and were initiated then required that detailed protocols be available to the personnel invol- ved. In particular, advice on tissue culture techniques, immunisation and animal handling, failures in growth and/or antibody secretion and the potential idiosyncrasies of monoclonal antibodies in assay and purifi- cation was sought. The book incorporates the accumulated experience of our own and other laboratories in this respect. Glasgow , June 1984 AILSAM . CAMPBELL Acknowledgements Many people have contributed to this book. In particular I should like to thank Marion Mc Cormack, Jacki Bennett and Souravi Ghosh who have been invaluable colleagues in the laboratory and Ian Ramsden and his staff in the Glasgow University Medical Illustration Unit who have been responsible for the illustrations. Janet Jones not only wrote Appendix I but taught us most of the techniques described in it. I am also indebted to Caroline Addey, Flora Campbell, Magnus Campbell, John Coggins, Dorothy Crawford, Bertil Damato, Morag Davidson, Jim Dunn, Christopher Edwards, Malcolm Ferguson Smith, Frank Fleming, Robin Fraser, Angus Munro, Rosalind Quinn, Celia Ross, Asaad Shallal, Martin Smellie, Elizabeth Smith, Alan Williamson and Veronica van Heyningen for information, support or advice. In addition, I thank the Glasgow University Biochemistry secretariat for help in word processing. Abbreviations EB Epstein Barr ELISA Enzyme linked immunosorbent assay G banding Staining of chromosomes with Giemsa stain HAT Hypoxanthine, aminopterin and thymidine HT Hypoxanthine and thymidine IRMA Immunoradiometric assay PAGE Polyacrylamide gel electrophoresis PEG Polyethylene glycol SDS Sodium dodecyl sulphate vii This Page Intentionally Left Blank Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . V Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . vii Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . vii Chapter I . General properties and applications of monoclonal an tibodies . . . . . . . . . . . . . . . . . . . . . . i 1.1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.2. Comparison of monoclonal antibodies and conventional antiserum . 4 1.2.1. Specificity: advantages . . . . . . . . . . . . . . . . . . . 5 1.2.2. Specificity: disadvantages . . . . . . . . . . . . . . . . . 8 1.2.3. Affinity: advantages . . . . . . . . . . . . . . . . . . . . 11 1.2.4. Affinity: disadvantages . . . . . . . . . . . . . . . . . . 11 1.2.5. Affinity and specificity: interrelationships . . . . . . . . . . 13 I .2. 6. Standardisation . . . . . . . . . . . . . . . . . . . . . . 12 . . . . . . . . . . . . . . . . . . . . . 1.2.7. Yield and purity 14 . . . . . . . . . . . . . . . . . 1.2.8. Purification requirements 14 1.2.9. Selection of immunoglobulin isotype . . . . . . . . . . . . 15 1.2.10. cost . . . . . . . . . . . . . . . . . . . . . . . . . . 16 1.3. Application of monoclonal antibodies . . . . . . . . . . . . . . 17 1.3.1. Diagnostic uses . . . . . . . . . . . . . . . . . . . . . . 17 1.3.2. Therapeutic uses . . . . . . . . . . . . . . . . . . . . . 20 1.3.3. Preparative uses . . . . . . . . . . . . . . . . . . . . . 25 . . . . . . . . . . . . . . . . . . . . . . 1.3.4. Basic research 29 1.3.5. T cell hybridomas . . . . . . . . . . . . . . . . . . . . . 30 Chapter 2 . Assay techniques . . . . . . . . . . . . . . . . . . 33 2.1. General assay requirements . . . . . . . . . . . . . . . . . . . 33 2.2. Theoretical considerations . . . . . . . . . . . . . . . . . . . . 33. ix
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