Analytical Characterization of Biotherapeutics

The definitive guide to the myriad analytical techniques available to scientists involved in biotherapeutics research
 
Analytical Characterization of Biotherapeutics covers all current and emerging analytical tools and techniques used for the characterization of therapeutic proteins and antigen reagents. From basic recombinant antigen and antibody characterization, to complex analyses for increasingly complex molecular designs, the book explores the history of the analysis techniques and offers valuable insights into the most important emerging analytical solutions. In addition, it frames critical questions warranting attention in the design and delivery of a therapeutic protein, exposes analytical challenges that may occur when characterizing these molecules, and presents a number of tested solutions.
 
The first single-volume guide of its kind, Analytical Characterization of Biotherapeutics brings together contributions from scientists at the leading edge of biotherapeutics research and manufacturing. Key topics covered in-depth include the structural characterization of recombinant proteins and antibodies, antibody de novo sequencing, characterization of antibody drug conjugates, characterization of bi-specific or other hybrid molecules, characterization of manufacturing host-cell contaminant proteins, analytical tools for biologics molecular assessment, and more.
* Each chapter is written by a recognized expert or experts in their field who discuss current and cutting edge approaches to fully characterizing biotherapeutic proteins and antigen reagents
* Covers the full range of characterization strategies for large molecule based therapeutics
* Provides an up-to-date account of the latest approaches used for large molecule characterization
* Chapters cover the background needed to understand the challenges at hand, solutions to characterize these large molecules, and a summary of emerging options for analytical characterization
 
Analytical Characterization of Biotherapeutics is an up-to-date resource for analytical scientists, biologists, and mass spectrometrists involved in the analysis of biomolecules, as well as scientists employed in the pharmaceuticals and biotechnology industries. Graduate students in biology and analytical science, and their instructors will find it to be fascinating and instructive supplementary reading.

List of contents

List of Contributors xv
 
1 Introduction to Biotherapeutics 1
Jennie R. Lill
 
1.1 Introduction 1
 
1.2 Types of Biotherapeutics and Manufacturing Systems 2
 
1.3 Types of Analyses Performed 5
 
1.4 Future perspectives 6
 
Acknowledgments 11
 
References 11
 
2 Mass Spectrometric Characterization of Recombinant Proteins 15
Corey E. Bakalarski, Wendy Sandoval, and Jennie R. Lill
 
2.1 Introduction 16
 
2.1.1 Ionization 16
 
2.1.1.1 Matrix Assisted Laser Desorption Ionization 17
 
2.1.1.2 Electrospray Ionization 19
 
2.1.2 Mass Analyzers for Intact Molecular Weight Measurement of Biotherapeutics 20
 
2.1.2.1 Time of Flight and Quadrupole Time of Flight Mass Spectrometers 20
 
2.1.2.2 High?]Resolution Intact Mass Measurement and Native MS 21
 
2.1.2.3 Ion Mobility Spectrometry 22
 
2.1.3 Software for the Analysis of Intact Molecular Weight Measurements 24
 
2.1.4 Separation Devices for the Characterization of Biotherapeutics 25
 
2.1.4.1 High?]performance Liquid Chromatography 25
 
2.1.4.2 Capillary Electrophoresis 26
 
2.1.4.3 Microfluidic Chromatographic Devices 28
 
2.2 Peptide Mass Fingerprinting 29
 
2.3 Tandem Mass Spectrometric Characterization of Biomolecules 30
 
2.3.1 Bottom-Up MS 33
 
2.3.2 Proteoinformatic Analysis of Bottom-Up Proteomic Data Sets 34
 
2.3.3 Top-Down MS 36
 
2.4 Conclusions and Perspectives 37
 
References 37
 
3 Characterizing the Termini of Recombinant Proteins 43
Nestor Solis and Christopher M. Overall
 
3.1 Introduction 44
 
3.2 Gel Electrophoresis and Edman Sequencing 46
 
3.3 Mass Spectrometric Approaches for Characterizing True Starts of Proteins 49
 
3.3.1 Top-Down Approaches 49
 
3.3.2 Current Caveats in Mass Spectrometric Identification of Protein Termini 54
 
3.3.3 Bottom-up Approaches for Identification of N- and C-Terminal Peptides 55
 
3.3.4 Amino Terminal Orientated Mass Spectrometry 56
 
3.3.5 Determining the True Start of Proteins from ATOMS LC-MS/MS Data 61
 
3.4 Conclusions 64
 
References 66
 
4 Assessing Activity and Conformation of Recombinant Proteins 73
Diego Ellerman, Till Maurer, and Justin M. Scheer
 
4.1 Introduction 74
 
4.2 Circular Dichroism 75
 
4.2.1 Applications of CD 77
 
4.2.1.1 Thermal Stability Analysis 77
 
4.2.1.2 Characterization of the Effect of PEGylation 77
 
4.2.1.3 Formulation and Stability Studies 77
 
4.2.1.4 Analysis of Biosimilars 78
 
4.2.2 Technical Improvements 78
 
4.3 DSC and Isothermal Titration Calorimetry 79
 
4.3.1 Use of DSC and ITC in Therapeutics Discovery 80
 
4.3.2 Protein Conjugation 82
 
4.3.3 Formulation and Stability 82
 
4.3.4 Analysis of Biosimilars 83
 
4.4 Hydrogen- Deuterium Exchange-Mass Spectrometry 85
 
4.4.1 Applications of HDX 86
 
4.4.1.1 Ligand-induced Conformational Changes and Mapping Interaction Sites 86
 
4.4.1.2 Applications in Protein Engineering 86
 
4.4.1.3 Comparability and Biosimilar Studies 88
 
4.4.1.4 Formulation and Aggregation Analysis 89
 
4.4.2 Technical Improvements and Challenges 89
 
4.5 Nuclear Magnetic Resonance 90
 
4.5.1 Applications of NMR 92
 
4.5.1.1 Flexible Proteins 92
 
4.5.1.2 Mapping Protein-Protein Interactions 93
 
4.5.1.3 Epitope Mapping 94
 
4.5.1.4 Protein Dynamics 94
 
4.5.1.5 Protein Conjugates and Complexes 94
 
4.5.1.6 Posttranslational

About the author

JENNIE R. LILL, PhD is Director of Proteomics at Genentech, where she is responsible for the management of Genentech's Microchemistry, Proteomics & Lipidomics Department. Dr. Lill has been involved in the experimental design and technological innovation of biological mass spectrometry for more than two decades. WENDY SANDOVAL leads the Applied Proteomics group at Genentech where she is responsible for the management and oversight of native mass spectrometry, antibody characterization, lipidomics and the core mass spectrometry group.

Summary

The definitive guide to the myriad analytical techniques available to scientists involved in biotherapeutics research Analytical Characterization of Biotherapeutics covers all current and emerging analytical tools and techniques used for the characterization of therapeutic proteins and antigen reagents.

Product details

• Authors: Jennie R. Lill, Jennie R. Sandoval Lill, JR Lill, Wendy Sandoval
• Assisted by: Jennie R. Lill (Editor), Lill Jennie R. (Editor), Jennie R Lill (Editor), Sandoval (Editor), Wendy Sandoval (Editor), Sandoval Wendy (Editor)
• Publisher: Wiley, John and Sons Ltd
• Languages: English
• Product format: Hardback
• Released: 03.10.2017
• EAN: 9781119053101
• ISBN: 978-1-119-05310-1
• No. of pages: 368
• Subjects: Natural sciences, medicine, IT, technology > Chemistry > Theoretical chemistry; Non-fiction book > Nature, technology > Natural science; Chemie, Biotechnologie, Massenspektrometrie, Life Sciences, Biowissenschaften, biotechnology, chemistry, mass spectrometry, Biotechnologie i. d. Biowissenschaften, Pharmaceutical & Medicinal Chemistry, Pharmazeutische u. Medizinische Chemie