Biophysical Characterization of Proteins in Developing Biopharmaceuticals

Damian J. Houde, Steven A. Berkowitz

• Publisher: Elsevier
• Year: 2014
• eBook ISBN: 9780444595904
• Topic: Biological Sciences
• Subtopic: Biochemistry
• Index: Biological Sciences

Biophysical Characterization of Proteins in Developing Biopharmaceuticals is concerned with the analysis and characterization of the higher-order structure (HOS) or conformation of protein based drugs. Starting from the very basics of protein structure this book takes the reader on a journey on how to best achieve this goal using the key relevant and practical methods commonly employed in the biopharmaceutical industry today as well as up and coming promising methods that are now gaining increasing attention. As a general resource guide this book has been written with the intent to help today's industrial scientists working in the biopharmaceutical industry or the scientists of tomorrow who are planning a career in this industry on how to successfully implement these biophysical methodologies. In so doing a keen focus is placed on understanding the capability of these methodologies in terms of what information they can deliver. Aspects of how to best acquire this biophysical information on these very complex drug molecules, while avoiding potential pitfalls, in order to make concise, well informed productive decisions about their development are key points that are also covered. - Presents the reader with a clear understanding of the real world issues and challenges in using these methods. - Highlights the capabilities and limitations of each method. - Discusses how to best analyze the data generated from these methods. - Points out what one needs to look for to avoid making faulty conclusions and mistakes. - In total it provides a check list or road map that empowers the industrial scientists as to what they need to be concerned with in order to effectively do their part in successfully developing these new drugs in an efficient and cost effective manner.

II

The Selected Biophysical Tools in the Biopharmaceutical Industry

Outline

Chapter 4. An Introduction and Hierarchical Organization of the Biophysical Tool in Section II

Chapter 5. The Value of UV, Fluorescence, and FTIR Spectroscopy in Biopharmaceutical Development

Chapter 6. Circular Dichroism Spectroscopy for Protein Characterization: Biopharmaceutical Applications

Chapter 7. Size-Exclusion Chromatograph (SEC) in Biopharmaceutical Process Development

Chapter 8. Scattering Techniques for the Characterization of Biopharmaceuticals

Chapter 9. Characterizing Biopharmaceuticals using Analytical Ultracentrifugation

Chapter 10. Subvisible and Visible Particle Analysis in Biopharmaceutical Research and Development

Chapter 11. Differential Scanning Calorimetry in the Biopharmaceutical Sciences

Chapter 12. Biophysical Mass Spectrometry for Biopharmaceutical Process Development: Focus on Hydrogen/Deuterium Exchange

Chapter 13. One- and Two-Dimensional NMR Techniques for Biopharmaceuticals∗

Chapter 4

An Introduction and Hierarchical Organization of the Biophysical Tool in Section II

Damian J. Houde, and Steven A. Berkowitz Department of Protein Pharmaceutical Development, Biogen Idec, Inc., Cambridge, MA, USA

Abstract

This chapter provides a brief introduction and hierarchical overview of the biophysical tools to be discussed in Section II of this book. In choosing these tools, we have organized them into a hierarchy of four separate levels (i.e., four tiers). The key criteria for this organization, concerns the richness in information each tool supplies regarding the biophysical properties and higher order structure of a protein drug. In using this key criterion a number of factors were observed to trend with this organization, including: (1) ease to operate and use, (2) cost to implement, (3) required amount of time to upset and acquire data, and (4) level of expertise to process, analyze and interpret data. The biophysical tools at the lowest two tiers (i.e., tier 1 and 2) fall into a general grouping that constitutes the standard or mainstream package of commonly used biophysical tools employed by nearly all biopharmaceutical companies in developing protein drugs. While the biophysical tools in the two highest tiers (i.e., tier 3 and 4), comprise a second grouping of methods that correspond to the more advanced biophysical tools.

Keywords

Advance biophysical tools; Standard biophysical tools

4.1. Introduction

In Section I (Chapters 1–3) of this book, details concerning the structure of a protein drug and its various structural elements that contribute to a protein's higher order structure (HOS) were discussed (see Chapter 1). Next, the biophysical characterization of a protein's HOS and how their biophysical properties play an important role in its development into a drug was examined (see Chapter 2). This was then coupled with a brief and general look into the basic biophysical toolbox used to achieve this characterization (see Chapter 3). In proceeding into Section II, the stage is now set to look more carefully and critically into a number of different biophysical tools. In choosing the particular biophysical tools discussed in this section, we have organized and categorized them into four different tiers based on the corresponding increase in informational content that they can provide. Hence, biophysical tools in tier 1, relatively speaking, provide the lowest level of information while biophysical tools in tier 4 provide, again relatively speaking, the highest level of information. In carrying out this categorization, we begin to see other trends as one makes the transition from tier 1 to tier 4 tools. In general, methods derived from tier 1 tools tend to be much easier to use and operate, are less expensive and less costly to implement, require the least amount of time to acquire the data, and need the least training or expertise to process, analyze, and interpret data. However, in moving toward tier 4 tools, the attributes of the methods in each subsequent tier become more difficult to use and operate, are more expensive and more costly to implement, requires more time to acquire the data and more training or expertise to process, analyze, and interpret data.

In creating these four tiers we have further arranged them to form two major groupings as shown in Figure 4.1. In the first grouping, only tier 1 and 2 tools have been included. Biophysical tools in this grouping consist of what we call the “standard” biophysical tools. These tools generally correspond to the most applicable and dominant biophysical tools used by many biopharmaceutical companies in characterizing and developing biopharmaceuticals. Methods generated from these tools form the backbone of many biophysical studies carried out in the biopharmaceutical industry and are found in virtually all regulatory filings.

FIGURE 4.1 Hierarchy of the biophysical tools in Section II. The biophysical tools that we have chosen to discuss in detail in Section II have been arranged into four tiers. In general, tier 1 tools correspond to those with low resolution, whereas tier 4 tools correspond to those capable of achieving high resolution. The biophysical tools in tier 1 and 2 have then been placed into a general grouping that we believe finds widest use throughout the biopharmaceutical industry as the “standard” biophysical tools. The biophysical tools in tier 3 and 4 have been placed into an advanced grouping, which represents the tools the industry is beginning to look at more seriously at for providing a more detailed picture of the protein drugs. Nuclear magnetic resonance (NMR), hydrogen deuterium exchange with mass spectroscopy detection MS (H/DX), small angle X-ray scattering (SAXS), differential scanning calorimetry (DSC), analytical ultracentrifugation (AUC), higher order structure (HOS), circular dichroism (CD), Fourier transform infrared spectroscopy (FTIR), size-exclusion chromatography (SEC), static light scattering/dynamic light scattering (SLS/DLS).

In the second grouping, only biophysical tools in tier 3 and 4 have been included. Tools in this grouping consist of what we call the more “advanced” biophysical tools. Although these tools are presently used in a limited capacity in the process development area, we anticipate their utility will expand in the coming years. Their ability to generate information with greater detail and higher resolution will help improve the industry's ability to characterize biopharmaceuticals to make better and sounder decisions concerning a range of critical evaluations. This should facilitate improvements in the overall biopharmaceutical development process by increasing the success rate of drug approvals, while at the same time reduce the cost and the time required in their development.

4.2. The Standard Class of Biophysical Tools Used in the Biopharmaceutical Industry

The biophysical tools listed in this category correspond to the standard set of tools used throughout the development process of most biopharmaceuticals. Nearly all of these tools will be employed in various phases of development work from comparability and stability studies to formulation development. In addition, they will likely be represented in characterization sections in investigational new drug and biological license application filings under the heading of biophysical or physicochemical characterization. In particular, size-exclusion chromatography (SEC) and particle analysis methods will, in most cases, be developed into lot release assays where they will be used to monitor aggregation. In the case of particle analysis, it is very likely this area will consist of more than one specific test (see Chapter 10). In addition to direct visual testing, obtaining information about subvisible and submicron particle analysis is becoming critical...

Table of contents

• Cover image
• Title page
• Table of Contents
• Copyright
• List of Contributors
• About the Editors
• Preface
• List of Abbreviations and Symbols
• I. Proteins and Biophysical Characterization in the Biopharmaceutical Industry
• II. The Selected Biophysical Tools in the Biopharmaceutical Industry
• III. Concluding Remarks on the Biophysical Characterization of Biopharmaceuticals
• Index