Copyright © 2005 by Marcel Dekker.

Copyright © 2005 by Marcel Dekker.

Although great care has been taken to provide accurate and current information, neither the author(s)
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Library of Congress Cataloging-in-Publication Data

Analytical techniques for biopharmaceutical development / Tim Wehr, Roberto Rodriguez-

Diaz, Stephen Tuck, editors.
p. ; cm.
Includes bibliographical references and index.
ISBN 0-8247-2667-7 (alk. paper)
1. Protein drugs--Analysis--Laboratory manuals.
[DNLM: 1. Pharmaceutical Preparations--analysis--Laboratory Manuals. 2.
Biopharmaceutics--methods--Laboratory Manuals. 3. Chromatography--methods--Laboratory
Manuals. 4. Electrophoresis--methods--Laboratory Manuals. 5. Spectrum Analysis--
methods--Laboratory Manuals. QV 25 A5338 2005] I. Wehr, Tim. II. Rodríguez-Díaz,
Roberto. III. Tuck, Stephen (Stephen F.)
RS431.P75A536 2005
615'.7--dc22
2004058292

ISBN: 0-8247-0706-0

This book is printed on acid-free paper.

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 Contents

About the Editors vii

Contributors ix

1. Analytical Techniques for Biopharmaceutical Development 1

Stephen Tuck

2. Introduction to the Development of Biopharmaceuticals 5

Roberto Rodriguez-Diaz

3. Protein Assay 13
Stephen Tuck and Rowena Ng

4. Use of Reversed-Phase Liquid Chromatography

in Biopharmaceutical Development 27
Tim Wehr

5. Practical Strategies for Protein Contaminant Detection

by High-Performance Ion-Exchange Chromatography 67
Pete Gagnon

6. Practical Strategies for Protein Contaminant Detection

by High-Performance Hydrophobic Interaction Chromatography 81
Pete Gagnon
v

Copyright © 2005 by Marcel Dekker.

vi Contents

7. Use of Size Exclusion Chromatography

in Biopharmaceutical Development 95
Tim Wehr and Roberto Rodriguez-Diaz

8. Slab Gel Electrophoresis for Protein Analysis 113

David E. Garfin

9. Capillary Electrophoresis of Biopharmaceutical Proteins 161

Roberto Rodriguez-Diaz, Stephen Tuck, Rowena Ng,
Fiona Haycock, Tim Wehr, and Mingde Zhu

10. Mass Spectrometry for Biopharmaceutical Development 227

Alain Balland and Claudia Jochheim

11. Analytical Techniques for Biopharmaceutical

Development — ELISA 279
Joanne Rose Layshock

12. Applications of NMR Spectroscopy in Biopharmaceutical

Product Development 305
Yung-Hsiang Kao, Ping Wong, and Martin Vanderlaan

13. Microcalorimetric Approaches to Biopharmaceutical Development 327

Richard L. Remmele, Jr.

14. Vibrational Spectroscopy in Bioprocess Monitoring 383

Emil W. Ciurczak

Copyright © 2005 by Marcel Dekker.

 About the Editors

Roberto Rodriguez-Diaz is Senior Scientist at Dynavax Technologies in Berke-

ley, California. He has extensive experience in product development in the bio-
pharmaceutical industry, and his research has focused on development of
analytical methodology ranging from determination of low molecular weight
reactants to analysis of protein-oligonucleotide conjugates. He holds a B.S. degree
from the University of Michoacan, Morelia, Mexico.

Tim Wehr is Staff Scientist at Bio-Rad Laboratories in Hercules, California. He

has more than 20 years of experience in biomolecule separations, including
development of HPLC and capillary electrophoresis methods and instrumentation
for separation of proteins, peptides, amino acids, and nucleic acids. He has also
worked on development and validation of LC-MS methods for small molecules
and biopharmaceuticals. He holds a B.S. degree from Whitman College, Walla
Walla, Washington, and earned his Ph.D. from Oregon State University in Corvallis.

Stephen Tuck is Vice President of Biopharmaceutical Development at Dynavax

Technologies in Berkeley, California. He has over 14 years of experience in
pharmaceutical chemistry. He was involved in the development of Fluad™ adju-
vated flu vaccine as well as various subunit vaccines, adjuvants, vaccine conju-
gates, and protein therapeutics. He earned his B.Sc. and Ph.D. degrees from
Imperial College, University of London, United Kingdom.

vii

Copyright © 2005 by Marcel Dekker.

 Contributors

Alain Balland Analytical Sciences, Amgen, Seattle, Washington

Emil W. Ciurczak Integrated Technical Solutions, Golden’s Bridge, New York

Pete Gagnon Validated Biosystems, Inc., Tucson, Arizona

David E. Garfin Garfin Consulting, Kensington, California

Fiona Haycock Dynavax Technologies, Berkeley, California

Claudia Jochheim Analytical Biochemistry, Corixa, Seattle, Washington

Yung-Hsiang Kao Genentech, Inc., South San Francisco, California

Joanne Rose Layshock Chiron Corporation, Emeryville, California

Rowena Ng Dynavax Technologies, Berkeley, California

Richard L. Remmele, Jr. Pharmaceutics, Amgen, Inc., Thousand Oaks,

California

Roberto Rodriguez-Diaz Dynavax Technologies, Berkeley, California

ix

Copyright © 2005 by Marcel Dekker.

x Contributors

Stephen Tuck Dynavax Technologies, Berkeley, California

Martin Vanderlaan Genentech, Inc., South San Francisco, California

Tim Wehr Bio-Rad Laboratories, Hercules, California

Ping Wong Genentech, Inc., South San Francisco, California

Mingde Zhu Bio-Rad Laboratories, Hercules, California

Copyright © 2005 by Marcel Dekker.