Hplc Of Peptides And Proteins

METHODS IN MOLECULAR BIOLOGY

TM TM

Volume 251

HPLC of Peptides and Proteins
Methods and Protocols
Edited by

Marie-Isabel Aguilar

M E T H O D S

I N

M O L E C U L A R

B I O L O G Y™

HPLC of Peptides and Proteins
Methods and Protocols

Edited by

Marie-Isabel Aguilar
Department of Biochemistry and Molecular Biology Monash University, Clayton, Victoria, Australia© 2004 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 www.humanapress.com All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. Methods in Molecular BiologyTM is atrademark of The Humana Press Inc. All authored papers, comments, opinions, conclusions, or recommendatoins are those of the author(s), and do not necessarily reflect the views of the publisher. This publication is printed on acid-free paper. ∞ ANSI Z39.48-1984 (American Standards Institute Permanence of Paper for Printed Library Materials.) Cover design by Patricia F. Cleary. Cover illustration:Cartoon depicting the selective retention of a peptide from a mixture of peptides and proteins. Illustration by Lee Tzong-Hsien, William Farrugia, and Marie-Isabel Aguilar. For additional copies, pricing for bulk purchases, and/or information about other Humana titles, contact Humana at the above address or at any of the following numbers: Tel.: 973-256-1699; Fax: 973-256-8341; E-mail:humana@humanapr.com; or visit our Website: www.humanapress.com Photocopy Authorization Policy: Authorization to photocopy items for internal or personal use, or the internal or personal use of specific clients, is granted by Humana Press Inc., provided that the base fee of US $25.00 per copy is paid directly to the Copyright Clearance Center at 222 Rosewood Drive, Danvers, MA 01923. For those organizationsthat have been granted a photocopy license from the CCC, a separate system of payment has been arranged and is acceptable to Humana Press Inc. The fee code for users of the Transactional Reporting Service is: [0-89603-977-3/04 $25.00 ]. Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1 ISSN: 1064-3745 E-ISBN: 1-59259-742-4 Library of Congress Cataloging-in-Publication Data HPLC ofpeptides and proteins : methods and protocols / edited by Marie-Isabel Aguilar. p. ; cm. -- (Methods in molecular biology ; 251) Includes bibliographical references and index. ISBN 0-89603-977-3 (alk. paper) 1. Peptides--Analysis. 2. Proteins--Analysis. 3. High performance liquid chromatography. [DNLM: 1. Peptides--chemistry. 2. Chromatography, High Pressure Liquid--methods. 3. Proteins-chemistry. QU 68H872 2004] I. Aguilar, Marie-Isabel. II. Methods in molecular biology (Totowa, N.J.); v. 251. QP552.P4H73 2004 572'.6536--dc22 2003017464

Preface
The introduction of high-performance liquid chromatography (HPLC) to the analysis of peptides and proteins some 25 years ago revolutionized the biological sciences by enabling the rapid and sensitive analysis of peptide and protein structure throughthe exquisite speed, sensitivity, and resolution that can be easily obtained. Today, HPLC in its various modes has become the pivotal technique in the characterization of peptides and proteins and currently plays a critical role in both our understanding of biological processes and in the development of peptide- and protein-based pharmaceuticals. The number of applications of HPLC in peptide andprotein purification continues to expand at an extremely rapid rate. Solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. HPLC techniques are also used extensively in the isolation and characterization of novel proteins that will become increasingly important in the postgenomic age....