The ELISA Guidebook
The Enzyme Linked Immunosorbent Assay (ELISA) continues, as predicted, to be the predominant assay used in biomedical and agricultural research. While little has changed in the principles of the ELISAs used since the publication of The ELIS
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Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
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METHODS
IN
MOLECULAR BIOLOGY™
The ELISA Guidebook Second Edition
by
John R. Crowther International Atomic Energy Agency, Animal Production & Health Section, Vienna, Austria
John R. Crowther International Atomic Energy Agency Animal Production & Health Section Wagramer Str. 5 1400 Vienna Austria
ISBN: 978-1-60327-253-7 e-ISBN: 978-1-60327-254-4 ISSN: 1064-3745 e-ISSN: 1940-6029 DOI: 10.1007/978-1-60327-254-4 Library of Congress Control Number: 2008940983 © Humana Press, a part of Springer Science+Business Media, LLC 2009 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013 USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. While the advice and information in this book are believed to be true and accurate at the date of going to press, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein. Cover illustration: Printed on acid-free paper springer.com
Preface There have been very few developments that markedly affect the need to greatly revise the text from the last version of this book. This is testament to the fact that heterogeneous enzyme-linked immunosorbent assays (ELISA) provide ideal systems for dealing with a wide range of studies in many biological areas. The main reason for this success is test flexibility, whereby reactants can be used in different combinations, either attached passively to a solid phase support or in the liquid phase. The exploitation of the ELISA has been increased through continued development of specifically produced reagents, for example, monoclonal and polyclonal antibodies and peptide antigens coupled with the improvement and expansion of commercial products such as enzyme-linked conjugates, substrates and chromogens, plastics technology and design of microwell plates, instrumentation advances and robotics. However, the principles of the ELISA remain the same. There has been some rearrangement of chapters plus addition of three new ones dealing with charting methods for assessing the indirect ELISA, ruggedness and robustness of tests-aspects of kit use and validation, and
