Analysis of Aggregates and Particles in Protein Pharmaceuticals

This book describes how to address the analysis of aggregates and particles in protein pharmaceuticals, provides a comprehensive overview of current methods and integrated approaches used to quantify and characterize aggregates and particles, and discusses regulatory requirements. Analytical methods covered in the book include separation, light scattering, microscopy, and spectroscopy.

Contributors ix

Preface xiii

Glossary xv

1 The Critical Need for Robust Assays for Quantitation and Characterization of Aggregates ofTherapeutic Proteins 1
John F. Carpenter, Barry Cherney, and Amy S. Rosenberg

I Methods Used for Detecting, Quantifying, and Sizing of Protein Aggregates and Particles 9

2 Separation-Based Analytical Methods for Measuring Protein Aggregation 11
Jun Liu, Barthélemy Demeule, and Steven J. Shire

3 Laser Light Scattering-Based Techniques Used for the Characterization of Protein Therapeutics 37
John den Engelsman, Fabian Kebbel, and Patrick Garidel

4 Online Detection Methods and Emerging Techniques for Soluble Aggregates in ProteinPharmaceuticals 61
Tapan K. Das

5 Analytical Methods to Measure Subvisible Particulates 85
Shawn Cao, Linda Narhi, Yijia Jiang, and Rahul S. Rajan

6 Detection of Visible Particles in Parenteral Products 117
Ronald Smulders, Hans Vos, and Hanns-Christian Mahler

7 Characterization of Aggregates and Particles Using Emerging Techniques 133
Hui Zhao, Manuel Diez, Atanas Koulov, Mariola Bozova, Markus Bluemel, and Kurt Forrer

II Methods Used to Characterize Protein Aggregates and Particles 169

8 Ultraviolet Absorption Spectroscopy 171
Reza Esfandiary and Charles Russell Middaugh

9 Fluorescence Spectroscopy to Characterize Protein Aggregates and Particles 201
Robert A. Poole, Andrea Hawe, Wim Jiskoot, and Kevin Braeckmans

10 Infrared Spectroscopy to Characterize Protein Aggregates 227
Marco van de Weert and Lene Jørgensen

11 Raman Microscopy for Characterization of Particles 249
Stefan Fischer, Oliver Valet, and Markus Lankers

12 Microscopic Methods for Particle Characterization in Protein Pharmaceuticals 269
Patrick Garidel, Andrea Herre, and Werner Kliche

III Integrated Approaches to Protein Aggregation and Particles 303

13 Comparison of Methods for Soluble Aggregate Detection and Size Characterization 305
John S. Philo

14 Protein Purification and Its Relation to Protein Aggregation and Particles 335
Roberto Falkenstein, Stefan Hepbildikler, Wolfgang Kuhne, Thorsten Lemm, Hans Rogl, Eva Rosenberg, Gerhard Winter, Frank Zettl, and Ralf Zippelius

15 Formulation Development and Its Relation to Protein Aggregation and Particles 369
Miriam Printz and Wolfgang Friess

Index 389

HANNS-CHRISTIAN MAHLER is Head of Pharmaceutical and Device Development, Pharma Technical Development Biologics Europe at F. Hoffmann-La Roche Ltd. He is also Lecturer at the Institute for Pharmaceutical Technology at the University of Frankfurt/Main, Germany.

WIM JISKOOT is Professor and Head of the Biologics Formulation Group in the Leiden/Amsterdam Center for Drug Research (LACDR) at Leiden University.

Enables researchers to better detect, measure, and characterize aggregates and particles during protein drug development

Featuring contributions from leading pharmaceutical investigators around the world, this text enables researchers to choose the most appropriate analytical methods in order to detect and characterize the full spectrum of protein aggregates and particles that can be present in a protein pharmaceutical. Moreover, the authors provide detailed examples of how these methods can be applied during both process and formulation development, helping researchers better manage the problem of protein instability as they seek to develop protein drugs.

The text begins with an introduction to aggregates and particles, focusing on their potential immunogenicity. Next, it covers:

• Methods to detect and measure soluble aggregates based on separation, light scattering, and new technology

• Methods to detect and measure insoluble aggregates and both sub-visible and visible particles

• Spectroscopic techniques such as UV/VIS, fluorescence, infrared and Raman spectroscopy, and microscopy to characterize aggregates and particles, elucidate their structures, and identify proteinaceous and non-proteinaceous particles

• Comparative advantages and disadvantages of the methods for protein aggregate analysis

• Approaches for managing aggregates and particles found during protein purification and formulation development

Throughout the text, case studies help researchers deal with the challenges that arise in detecting and analyzing protein aggregates and particles.

Protein pharmaceuticals are increasingly used to treat life-threatening and chronic diseases, including cancer, viral infections, metabolic disorders, and central nervous system diseases. With Analysis of Aggregates and Particles in Protein Pharmaceuticals as their guide, pharmaceutical researchers now have an important tool in overcoming the challenging problem of aggregates and particles in protein drug development.