1. No category

F
BIOPHYSICAL
CHARACTERIZATION
OF PROTEINS IN
DEVELOPING
BIOPHARMACEUTICALS
Edited by
J.
DAMIAN
HouDE
Biogen Idee Inc., Department of Protein Pharmaceutical Development, Cambridge, MA
STEVEN
A.
BERKOWITZ
Biogen Idee Inc., Department of Protein Pharmaceutical Development, Cambridge, MA
ELSEVIER
AMSTERDAM • BOSTON • HEIDELBERG • LONDON • NEW YORK • OXFORD
PARJS • SAN DIEGO • SAN FRANCISCO • SINGAPORE • SYDNEY • TOKYO
p
Contents
,
'
1ents
:y, can
the
List of Contributors ix
About the Editors xi
Preface xiii
List of Abbreviations and Symbols
2. Biophysical Characterization and Its Role
in the Biopharmaceutical
Industry 23
DAMIAN
xvii
I
ume
ility,
PROTEINS AND BIOPHYSICAL
CHARACTERIZATION IN
THE BIOPHARMACEUTICAL
INDUSTRY
3. Biopharmaceutical Industry's Biophysical
Toolbox 49
1. The Complexity of Protein Structure and
the Challenges it Poses in Developing
Biopharmaceuticals 1
STEVEN A. BERKOWITZ, DAMIAN
J.
HOUDE, SlEVEN A. BERKOWITZ
2.1 Drug Development Process 23
2.2 Protein Drugs (Biopharmaceuticals) 25
2.3 The Role of Biophysical Characterization in
Biopharmaceutical Drug Development 27
2.4 The Challenges in Conducting Biophysical
Measurements to Detect Changes in a Protein
Drug's HOS 39
2.5 Regulatory Needs and Considerations 42
References 43
Further Reading 47
ience
l.
J.
DAMIAN
J.
HOUDE, STEVEN A. BERKOWITZ
3.1 Attributes of a Single Biophysical Tool to
Characterize and Detect Changes in the
Higher Order Structure of a
Biopharmaceutical 49
3.2 Studying the Biophysical Properties of a
Biopharmaceutical as an Indirect
Approach for Characterizing Changes in its
HOS 51
3.3 General Considerations in Analyzing the
Biophysical Properties of
Biopharmaceuticals 54
3.4 The Utility of Using Stress to Monitor Changes
in the HOS Profile of a Protein Drug 58
3.5 Present Biophysical Toolbox 59
3.6 Conclusion 74
References 74
Further Reading 78
HOUDE
1.1 The Basics of Protein Higher-Order
Structure (HOS) 1
1.2 The Search for How Proteins Attain
Their Correct HOS: The Protein Folding
Problem 8
1.3 Surprises in the World of Protein Folding:
Intrinsically Disordered or Unstructured
Proteins (An Apparent Challenge to
the Protein Structure-Function
Paradigm) 12
1.4 Proteins and the Biopharmaceutical
Industry: Problems and Challenges 12
1.5 Conclusion 17
References 18
Further Reading 21
v
--------------..
vi
CONTENTS
II
THE SELECTED
BIOPHYSICAL TOOLS IN THE
BIOPHARMACEUTICAL
INDUSTRY
4. An Introduction and Hierarchical
Organization of the Biophysical Tool in
Section II 79
4.1 Introduction 79
4.2 The Standard Class of Biophysical Tools Used
in the Biopharmaceutical Industry 81
4.3 The Advanced Class of Biophysical Tools
Used in the Biopharmaceutical Industry
82
84
5. The Value of UV, Fluorescence, and
FTIR Spectroscopy in
Biopharmaceutical Development 87
MARK BRADER
5.1 Introduction 87
5.2 The Origins of Electronic Absorption,
Fluorescence, and FT-IR Spectroscopy 88
5.3 Conformational Analysis of Proteins in
Solution 92
5.4 Optical Spectroscopy and Product
Comparability 95
5.5 Optical Spectroscopy and High-throughput
Methods 99
5.6 Solid-State Measurements 102
5.7 Conclusions 104
References 105
6. Circu l.ar Di nroi m Spectro c py for
Pr tein Chara terizati n:
Bi ph.armac uti a l Applications 109
A.]. MILES, B.A. WALLACE
6.1 Introduction 109
6.2 Instrumentation 114
Data Generated 115
Guid o olle ting c>Od Daro 1 L6
Da. Pro c · ing <md Analys
127
R le in the Re earch Industry 1 0
Techn logy Availability 131
6.8 Fucurc D .vel pmcnts 133
Acknowledgments 134
References 134
Further Reading 136
7. Size-Exclusion Chromatograph (SEC) in
Biopharmaceutical Process
Development 139
DAMIAN). HOUDE, STEVEN A. fiERKOWITZ
4.4 An Overview of Section II
References 84
Further Reading 85
6.3
6.4
6.
6.6
6.7
STEVEN A. BERKOWITZ, DAMIAN). HOUDE
7.1 Introduction 139
7.2 Basic Theory of Normal or Ideal
SEC 140
7.3 Maximizing Sec Separation By Enhancing
The Usage Of Pore Volume and Pore
Structure 144
7.4 Characteristics of Pore Structure 146
7.5 Nonideal SEC Chromatography 147
7.6 Assessing and Maintaining an Optimum SEC
Chromatography Method 151
7. 7 Detectors 153
7.8 Multidetector SEC 160
7.9 Aggregation 164
7.10 Technology Advances 165
7.11 Conclusion 165
References 166
8. Scattering Techniques for the
Characterization of
Biopharmaceuticals 171
LEE MAKOWSKI, STEVEN A. BERKOWITZ,
DAMIAN ]. HOUDE
8.1 Introduction
171
8.2 Intensity- and Time-Dependent Light
Scattering 172
8.3 General Comment Concerning SLS and
DLS 185
8.4 The '' u t Probl m" in SLS and DLS 186
8.5 X-Ray cattering: haracterization of Proteins
in olution U. ing mall-Angle X-Ray
catterin
187
References 206
1
vii
CONTENTS
1ta
116
127
I
130
9. Characterizing Biopharmaceuticals
using Analytical
Ultracentrifugation 211
STEVEN A. BERKOWITZ, JOHN S. PHILO
ph (SEC) in
rocess
39
<\Nj. HOUDE
al
Enhancing
d Pore
146
147
:Jptimum SEC
:e
9.1 Introduction 211
9.2 Unique Features of the Analytical
Ultracentrifuge That Make It Different from
Other Centrifuges 212
9.3 Theory 213
9.4 Utility of AUC in the Biopharmaceutical
Industry 217
9.5 Boundary SV-AUC 219
9.6 Band SV-AUC 239
9.7 Sedimentation Equilibrium, SE-AUC 241
9.8 Density-Gradient SE-AUC 243
9.9 AUC Detectors 245
9.10 Miscellaneous Helpful Information About
Conducting AUC Experiments 252
9.11 Conclusion 254
References 255
Further Reading 259
y
10. Subvisible and Visible Particle Analysis
in Biopharmaceutical Research and
Development 261
ANDREA HA WE, SARAH ZOLLS, ANGELIKA
FREITAG, JOHN F. CARPENTER
e
ITZ,
10.1 Introduction 261
10.2 Overview of Analytical Methods 262
10.3 General Recommendations and Pitfalls for
Particle Analysis 279
10.4 Outlook and Conclusions 282
References 283
11. Differential Scanning Calorimetry in
the Biopharmaceutical Sciences 287
:._ight
;Ls and
DLS 186
m of Proteins
:-Ray
STEPHEN). DEMAREST, VERNA FRASCA
11.1
11.2
11.3
11.4
Background 287
DSC Instruments 293
Practical Considerations for DSC Use
Data Analysis 299
295
11.5 Applications of Solution DSC in
Biopharmaceutical Discovery and
Development 300
11.6 Applications of Solid-Sample DSC in
Biopharmaceutical Discovery and
Development 302
11.7 Conclusions 304
Acknowledgments 304
References 304
12. Biophysical Mass Spectrometry for
Biopharmaceutical Process
Development: Focus on Hydrogen/
Deuterium Exchange 307
GEORGE M. BOU-ASSAF, ALAN G. MARSHALL
12.1
12.2
12.3
12.4
12.5
12.6
Introduction 307
Synopsis of the Technique 311
Mechanism of Exchange 312
Advances in the Technique 314
Commercialization 322
Applications in the Biopharmaceutical
Industry 323
12.7 Future Perspective 330
Acknowledgments 331
References 331
13. One- and Two-Dimensional NMR
Techniques for
Biopharmaceuticals 341
YVES AUBIN, DARON I. FREEDBERG,
DA VJD A. KEIRE
13.1 Physical Basis of the Technique 341
13.2 The Appropriate Technique for a Particular
Problem 356
13.3 Method Requirements and
Performance 361
13.4 Data Processing (Procedures) 373
13.5 Role in Research vs Process
Development 375
13.6 Technology Update: Recent and Future
Advances and Unique Applications 377
13.7 References 378
Further reading 382
viii
C.'ONTENTS
III
CONCLUDING REMARKS
ON THE BIOPHYSICAL
CHARACTERIZATION
OF BIOPHARMACEUTICALS
14. Biophysical Characterization: An
Integral Part of the "Totality of the
Evidence" Concept 385
14.3 Bi physical Characterization in Developing
Prot in Biopharma eu ica! 388
14.4 Bu ilding a Biopharmaceutical's Biophysical
Ping rprint 389
14.5 Detecting Small Differences in
Biopharmaceuticals via Biophysical
Characterization Meas urements 393
14.6 Conclusion 395
References 395
Index
397
DAMIAN). HOUDE, STEVEN A. BERKOWITZ
14.1 Biopharmaceutical Development 385
14.2 An Introduction to the "Totality of the
Evidence" and Its More Global Meaning in
Develop ing Biopharmaceuticals 386
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