Monoclonal Antibody and Immunosensor Technology: The Production and Application of Rodent and Human Monoclonal Antibodies PDF
Preview Monoclonal Antibody and Immunosensor Technology: The Production and Application of Rodent and Human Monoclonal Antibodies
Monoclonal antibody and immunosensor technology The production and application of rodent and human monoclonal antibodies LABORATORY TECHNIQUES IN BIOCHEMISTRY AND MOLECULAR BIOLOGY Volume 23 Edited by P.C. van der VLIET - Department for Physiological Chemistry, University of Utrecht, Utrecht ELSEVIER AMSTERDAM. LONDON. NEW YORK . TOKYO MONOCLONAL ANTIBODY AND IMMUNOSENSOR TECHNOLOGY The production and application of rodent and human monoclonal antibodies Ailsa M. Campbell Department of Biochemistry, University of Glasgow, Glasgow GI2 SQQ, U.K. 1991 ELSEVIER AMSTERDAM. LONDON. NEW YORK . TOKYO 01991, Elsevier Science Publishers B. V.A ll rights reserved No part of this publication may be reproduced, stored in a retrieval system. or transmitted in any form or by any means, electronic. mechanical, photocopying, recording or other- wise, without the prior written permission of the Publisher, Elsevier Science Publishers B. V., Permissions Department, P.O. Box 521, I000 AN Amsterdam, The Netherlands. No responsibility is assumed by the Publisher for any injury andlor damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods. products. instructions or ideas contained in the material herein. Because of the rapid advances in the medical sciences, the Publisher recommends that in- dependent verijication of diagnoses and drug dosages should be made. Special regulations for readers in the U.S.A.: This publication has been registered with the Copyright Clearance Center, Inc. (CCC), Salem, Massachusetts. Information can be obtained from the CCC about conditions under which the photocopying of parts of this publication may be made in the U.S.A. All other copyright questions, includingphotocopy- ing outside ihe U.S.A.. should be referred to the Publisher. ISBN 0-444-81412-4 (pocket edition) ISBN 0-444-81413-2 (library edition) ISSN 0-7204-4200-1 (series) Published by: ELSEVIER SCIENCE PUBLISHERS B.V. P.O. BOX 21 1 1000 AE AMSTERDAM THE NETHERLANDS Sole distributors for the U.S.A. and Canada: ELSEVIER SCIENCE PUBLISHING COMPANY, INC. 655 AVENUE OF THE AMERICAS NEW YORK, NY 10010 U.S.A. Library of Congress Card No. 85-647011 Printed in the Netherlands on acid-free paper Preface This book is the successor to a book in the same series entitled Mono- clonal Antibody Technology, published in 1984. Even at that time it was apparent that unrealistic projections were being made for a useful but not omnipotent technology. The main problem is, and has always been, that it is possible to obtain invaluable monoclonal antibodies as reagents, and also to obtain useless ones. The skill lies in the selec- tion of high affinity Mabs by a method which is close to the final use and far too many research strategies have concentrated on quantity (numbers of positive clones) rather than quality. It has often been dis- tressing to see the technique abused by those who do not appreciate the importance of affinity and native antigen presentation. The basic method for Mab production remains unchanged and fu- sion and cloning methods are close to those used 10 years ago. The main changes relate to the methodology of selection and use. In the early 1980s people sought Mabs to simple soluble proteins and there is now a vast library of Mabs to abundant proteins such as lysosyme, all with no application. In contrast, there is an increasing interest in Mabs to cell membrane proteins and these are scarcely ever produced by the generation of Mabs to denatured proteins or peptides, since epitopes are usually conformational. Thus selection techniques have changed. Too many researchers fail to understand that a monoclonal anti- body is not necessarily monospecific. I have emphasised this point heavily in various sections of the book. A good Mab is usually mono- specific unless slammed up against the wall of an ELISA plate against molar amounts of irrelevant antigen in which case it may give a slight signal. A bad Mab will cross-react with a wide range of antigens. The methods of obtaining the former are detailed. V vi This book has a slight bias towards Mabs directed to supposed tu- mour antigens. I have been astonished by the frequency with which Mabs to uncharacterised antigens have been used, to predictably little effect in ‘tumor therapy’, especially as the adverse side effects of a mouse Mab are considerable (Section 1.9). Almost all these Mabs are to high density structural antigens which, in the light of modern un- derstanding of oncogene product function, are irrelevant to tumour progression. This tendency is unfortunate, since far better Mabs could be made in the light of modern knowledge of the nature of tumour progression. The book also covers the concepts behind the emerging biosensor techniques (Chapter 12). Frankly, having made my way through the mystiques of the systems, I was surprised by their primitive low detec- tion limits despite their cost and cumbersome nature. It would appear that there has been a breakdown in communications between Mab technologists and electrochemical or opto-electronic engineers where the latter group have assumed that antibodies have exquisite specifi- city and no cross-reactive potential and the former have failed to dis- abuse them of this fact. In terms of classic detection systems, for my own work, I am now proceeding to use enzyme-linked chemilumines- cent detection systems and all instant camera. I have also covered the gene cloning strategies for Mabs and again have found that many of the complex systems, once understood and applied, are unsuitable for Mab production. In particular, PCR gene- rated variable gene libraries (Section 9.6) are technically unsound for reasons stated in the text. However, other aspects of genetic manipu- lation involving PCR are very exciting and the prospect of immortal- king human Mab responses by PCR of early unstable, but high affini- ty clones seems to have immense potential. Tobacco plants and alfalfa looping moths notwithstanding, the best expression system is still a myeloma cell. vii I have included a section on Mab patents which I particularly en- joyed investigating. The fact that the original Kohler-Milstein fusion procedure was not patented means that no financial concern or com- pany promotes it. I should like to do so. It is still the best way of mak- ing a good Mab. Ailsa Campbell January 199 1. This Page Intentionally Left Blank Acknowledgements I should like to thank Celia Cannon, Pat Ferry, Souravi Ghosh, Lor- raine Haynes and Delyth Wong in particular. I should also like to acknowledge help from Munir Alam, Peggy Anderson, Rob Butcher, Flora Campbell, Magnus Campbell, Sue Christie, Bill Cushley, Bertil Damato, Chris Darnborough, Morag Davidson, Fiona Durie, Alan Hughes, Janet Jones, Moira McCann, Tom Mathieson and Ian Ramsden. ix
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