Physical Methods to Characterize Pharmaceutical Proteins PDF
Preview Physical Methods to Characterize Pharmaceutical Proteins
Physical Methods to Characterize Pharmaceutical Proteins Pharmaceutical Biotechnology Series Editor: Ronald T. Borchardt The University ofK ansas Lawrence, Kansas Volume 1 PROTEIN PHARMACOKINETICS AND METABOLISM Edited by Bobbe L. Ferraiolo, Marjorie A. Mohler, and Carol A. Gloff Volume 2 STABILITY OF PROTEIN PHARMACEUTICALS, Part A: Chemical and Physical Pathways of Protein Degradation Edited by Tim J. Ahem and Mark C. Manning Volume 3 STABILITY OF PROTEIN PHARMACEUTICALS, Part B: In Vivo Pathways of Degradation and Strategies for Protein Stabilization Edited by Tim J. Ahem and Mark C. Manning Volume 4 BIOLOGICAL BARRIERS TO PROTEIN DELIVERY Edited by Kenneth L. Audus and Thomas J. Raub Volume 5 STABILITY AND CHARACTERIZATION OF PROTEIN AND PEPTIDE DRUGS: Case Histories Edited by Y. John Wang and Rodney Pearlman Volume 6 VACCINE DESIGN: The Subunit and Adjuvant Approach Edited by Michael F. Powell and Mark J. Newman Volume 7 PHYSICAL METHODS TO CHARACTERIZE PHARMACEUTICAL PROTEINS Edited by James N. Herron, Wim Jiskoot, and Daan J. A. Crommelin Physical Methods to Characterize Pharmaceutical Proteins Edited by James N. Herron University of Utah Salt Lake City. Utah WimJiskoot National Institute ofP ublic Health and Environmental Protection Bilthoven. The Netherlands and Daan J. A. Crommelin Utrecht University. and Utrecht Institutefor Pharmaceutical Sciences Groningen-Utrecht Institutefor Drug Exploration Utrecht. The Netherlands Springer Science+Business Media, LLC Library of Congress Cataloging-in-Publication Data On file ISBN 978-1-4899-1081-3 ISBN 978-1-4899-1079-0 (eBook) DOI 10.1007/978-1-4899-1079-0 © Springer Science+Business Media New York 1995 Originally published by Plenum Press, New York in 1995 Softcover reprint of the hardcover 1st edition 1995 109 8 765 43 2 1 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 Contributors Michael Bloemendal • Department of Protein and Molecular Biology, Royal Free Hospital School of Medicine, London NW3 2PF, England E. A. Cooper • Department of Bioengineering, University of Utah, Salt Lake City, Utah 84112 Reinout J. Driebergen • Ares Serono, Chemin des Mines, Geneva, Switzer land Ernesto Freire • Department of Biology and Biocalorimetry Center, The Johns Hopkins University, Baltimore, Maryland 21218 Paul T. Hamilton • Becton Dickinson Research Center, Research Triangle Park, North Carolina 27709 James N. Herron • Department of Pharmaceutics and Pharmaceutical Chemis try, University of Utah, Salt Lake City, Utah 84112 Vladimir Hlady • Department of Bioengineering, University of Utah, Salt Lake City, Utah 84112 Joost J. M. Holthuis • OctoPlus b.v., 2300 AS Leiden, The Netherlands Wim Jiskoot • Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, Utah 84112; present address: Laboratory for Product and Process Development, National Institute of Public Health and Environmental Protection, BA Bilthoven, The Netherlands W. Curtis Johnson, Jr. • Department of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon 97331 K. Knutson • Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, Utah 84112 Mark C. Manning • School of Pharmacy, University of Colorado Health Sci ences Center, Denver, Colorado 80262 v vi Contributors James Matsuura • School of Pharmacy, University of Colorado Health Sci ences Center, Denver, Colorado 80262 Kenneth P. Murphy • Department of Biochemistry, University of Iowa, Iowa City, Iowa 52242 John J. Naleway • Marker Gene Technologies, University of Oregon, Eugene, Oregon 97403 Peter Roepstorjf • Department of Molecular Biology, Odense University, DK-5230 Odense M, Denmark Tom A. A. M. van de Goor • Eindhoven University of Technology, Laboratory of Instrumental Analysis, 5600 MB Eindhoven, The Netherlands; present address: Hewlett Packard Laboratories, Palo Alto, California 94303-0867 David G. Vander Velde • NMR Laboratory, University of Kansas, Lawrence, Kansas 66045 Preface Proteins are still gaining importance in the pharmaceutical world, where they are used to improve our arsenal of therapeutic drugs and vaccines and as diagnostic tools. Proteins are different from "traditional" low-molecular-weight drugs. As a group, they exhibit a number of biopharmaceutical and formulation problems. These problems have drawn considerable interest from both industrial and aca demic environments, forcing pharmaceutical scientists to explore a domain previ ously examined only by peptide and protein chemists. Biopharmaceutical aspects of proteins, e.g., low oral bioavailability, have been extensively investigated. Although all possible conventional routes of ad ministration have been examined for proteins, no real, generally applicable alter native to parenteral administration in order to achieve systemic effects has yet been discovered. Several of these biopharmaceutical options have been discussed in Volume 4 of this series, Biological Barriers to Protein Delivery. Proteins are composed of many amino acids, several of which are notorious for their chemical instability. Rational design of formulations that optimize the native structure and/or bioactivity of a protein is therefore of great importance when long shelf life is required, as it is for pharmaceutical products. This issue has also been examined in two prior volumes of this series: Volume 2: Stability of Protein Pharmaceuticals (Part A) and Volume 5: Stability and Characterization of Protein and Peptide Drugs. Of equal importance for the therapeutic or diagnostic success of proteins is their physical stability. The integrity of their secondary, tertiary, and quaternary structure should be guaranteed during their shelf life (e.g., liquid formulations) or upon administration to the patient (e.g., freeze-dried products). This means that techniques that provide information about various structural aspects of proteins have to be used. These aspects include, but are not limited to, three-dimensional structure, hydrodynamic properties, physicochemical behavior, kinetics, thermo dynamic properties, and dynamic behavior. Unfortunately, no one technique in the present arsenal of structural methods is able to provide 100% of this information. Therefore, a rational strategy is to vii viii Preface employ a concerted approach in which the protein is examined using several different structural techniques. The resulting information is cross-correlated to provide a more complete picture of the chemical and physical state and/or bio activity of the protein under different conditions. The most frequently used techniques for structural analysis of proteins are described in this volume. The impressive recent progress in all these techniques and expected future develop ments are also discussed. Several spectroscopic techniques can be employed for studying the structure and function of proteins. These include fluorescence spectroscopy (Chapter I), circular dichroism (Chapter 2), and infrared spectroscopy (Chapter 3). The first of these, fluorescence spectroscopy, exhibits a level of sensitivity (subnanomolar) unachievable by any other technique described in this volume. For that reason, it is especially well suited for protein function studies where information about ligand-receptor interactions (including antigen-antibody binding) and enzyme kinetics is required. Furthermore, recent advances in time-resolved fluorescence have made it possible to study dynamic processes that occur in proteins in the nanosecond to microsecond time scale. Circular dichroism (CD) is a spectroscopic technique that has long been part of the concerted approach. Different parts of the spectrum (far UV, near UVNIS, infrared) provide information on the secondary and tertiary structure of proteins. The degree of structural information is less detailed than with new nuclear magnetic resonance (NMR) or X-ray diffraction techniques, but CD has the advantage that relatively simple equipment is used, scans are rapidly obtained, and the (semi) empirical interpretation is not complicated. It has long been known that certain transitions in the infrared spectrum of a protein were due to the vibrations of the carbonyl and amide functions of the peptide bond, but only in the last decade have instrumentation and analytical techniques improved to the point where these transitions could be effectively used to predict the secondary structure of a protein. Thus, infrared spectroscopy, or more correctly, Fourier transform infrared (FTIR) spectroscopy, has become a viable technique for examining secondary structure. Although the information obtained is similar to that provided by CD spectroscopy, FTIR offers a greater flexibility in detection that facilitates measurements with specimens such as cells, crystals, tissue slices, and thin films, in addition to aqueous solutions of proteins. In the last decade, the role of mass spectrometry for protein characterization changed from only a marginal one to a core position. This dramatic change is mainly the result of the development of a number of new ionization methods. Potentials and limitations of these new mass spectrometry approaches and future developments are discussed in Chapter 4. As with mass spectrometry, the role of NMR spectroscopy in protein charac terization has changed dramatically in the last decade and now rivals that of X-ray diffraction, at least in the case of small-to medium-sized proteins (10-50 kDa). This has largely been due to three developments: the availability of high- Preface ix resolution spectrometers (both high magnetic field and high frequency), the effective usage of spin-coupling relaxation times to provide two-dimensional data, and the labeling of proteins with isotopes such as 13C and 15N to provide data of even higher dimensionality (three- and four-dimensional) than is available with standard proton (IH) NMR experiments. These developments and their applica tion to proteins of pharmaceutical interest, are discussed in Chapter 5. X-ray diffraction has been and continues to be the definitive method for determining the three-dimensional structure of a protein. However, the technique is usually performed by specialists, and four excellent works reviewing the state of the art in X-ray diffraction analysis of proteins appeared recently. Besides, X-ray crystallography is not a technique that can be easily used in quality control protocols of pharmaceutical proteins because of the problems encountered with crystallization. Therefore, we have not included a chapter on X-ray diffraction in the present volume and refer interested readers to recent works by Ducruix and Giege (1992), Rhodes (1993), McRee (1994), and Drenth (1994). Thermodynamic parameters provide valuable information about stability, and differential scanning calorimetry has been used for many years in the charac terization of small molecules. However, it has only been in the last 5 to 10 years that microcalorimetry instrumentation has progressed to the point where similar measurements could be obtained for macromolecules. Recent developments in this field and their application to both protein folding and protein-ligand inter actions are discussed in Chapter 6. Chromatographic approaches have been the standard-bearers of the con certed approach mentioned above. As discussed in Chapter 7, several fundamen tally different chromatographic separation approaches, each with its own pros and cons, have been developed over the years. Reversed-phase chromatography, hydrophobic interaction chromatography, ion-exchange chromatography, size exclusion chromatography, affinity and immunoaffinity chromatography, and perfusion chromatography all provide characteristic information about the peptide or protein involved. This tendency to diverge has made chromatography a highly flexible tool for monitoring different aspects of proteins. Equally important for optimization of the separation process is the further improvement of detectors. The reader is therefore informed about, for example, "the state of the art" in hyphenated high-performance liquid chromatography-mass spectrometry config urations (Chapters 4 and 7). A new branch on the tree of protein characterization methodologies is capillary electrophoresis (Chapter 8). The potential of capillary electrophoresis for the characterization of peptides and proteins has not yet been fully explored, but it is clear that upon "maturation" of the technique in the future, capillary electrophoresis will be recognized as extremely important because it provides data complementary to other approaches. We would be remiss not to include a chapter about the impact of molecular biology on the characterization of pharmaceutical proteins. First, a number of such
The list of books you might like
