Ribosome Display and Related Technologies Methods and Protocols
Display technologies have become a very powerful way of generating therapeutic lead molecules and specific reagents for increasing our understanding of biology; however, despite being first described shortly after phage display, the use of ribosome displa
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IN
MOLECULAR BIOLOGY™
Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: http://www.springer.com/series/7651
Ribosome Display and Related Technologies Methods and Protocols Edited by
Julie A. Douthwaite and Ronald H. Jackson Department of Antibody Discovery and Protein Engineering, MedImmune Limited, Cambridge, UK
Editors Julie A. Douthwaite, Ph.D Department of Antibody Discovery and Protein Engineering MedImmune Limited Cambridge, UK [email protected]
Ronald H. Jackson, Ph.D Department of Antibody Discovery and Protein Engineering MedImmune Limited Cambridge, UK [email protected]
ISSN 1064-3745 e-ISSN 1940-6029 ISBN 978-1-61779-378-3 e-ISBN 978-1-61779-379-0 DOI 10.1007/978-1-61779-379-0 Springer New York Dordrecht Heidelberg London Library of Congress Control Number: 2011940807 © Springer Science+Business Media, LLC 2012 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. Printed on acid-free paper Humana Press is part of Springer Science+Business Media (www.springer.com)
Preface Over the last 20 years, display technologies have become a very powerful way of generating therapeutic lead molecules and specific reagents for increasing our understanding of biology. Despite first being described shortly after phage display, the use of ribosome display and related methods has been much less widespread. Since this is in part due to the complexity of the methods, it is our hope that the availability of this volume of Methods in Molecular Biology™ will allow their extended use. The protocols described range from wellestablished methods that have been used for a decade to generate high affinity antibodies which are in the clinic to methods that are more at their early stages of application, such as display of peptides incorporating noncanonical amino acids. At the core of ribosome display and related technologies, such as mRNA display, “in vitro virus,” and cDNA display, is the in vitro generation of a library of diverse molecules in which a peptide or protein is associated with the nucleic acid encoding it. From the large libraries of over 1012 variants that can be made by these purely in vitro methods, molecules with desired properties can be selected by binding to a partner molecule. In all of these methods, the displayed prot
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