Analytical characterization of biotherapeutics. (2017)

• Record Type: Book
• Further Information: Note: Edited by Jennie R. Lill, Wendy Sandoval.
• Editors: Lill, Jennie R; Sandoval, Wendy N
• 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 Modifications 95 4.5.1.7 Biosimilars 95 4.6 Concluding Remarks 96 References 98 5 Structural Characterization of Recombinant Proteins and Antibodies 111; Paola Di Lello and Patrick Lupardus 5.1 Introduction 112 5.2 Antigens, Epitopes, and Paratopes 113 5.2.1 Rationale for Structural Characterization of Epitopes 113 5.3 Choice of Analytical Method for Epitope Mapping 117 5.3.1 EM for Epitope Analysis 117 5.3.2 Epitope and Paratope Mapping by NMR 118 5.3.2.1 Epitope/Paratope Mapping by Chemical Shift Perturbations 119 5.3.2.2 Final Considerations 122 5.3.3 Epitope Mapping by X-ray Crystallography 122 5.4 Recombinant Antigen Generation 123 5.4.1 E. coli Expression of Antigens 124 5.4.2 Insect Cell Expression of Antigens 125 5.4.3 Mammalian Expression of Antigens 126 5.5 N-linked Glycosylation 127 5.5.1 E. coli Expression to Remove Glycosylation as a Factor 128 5.5.2 Manipulating N-linked Glycans on Antigens 128 5.6 Antibody Generation for Crystallography 129 5.7 Crystallization of Antibody/Antigen Complexes 130 5.8 Conclusion 131 References 131 6 Antibody de novo Sequencing 139; Natalie Castellana and Adrian Guthals 6.1 Introduction 139 6.2 Technical Details on Antibody de novo Sequencing 141 6.2.1 Achieving Complete Protein Coverage 141 6.2.2 Achieving High Sequencing Accuracy 142 6.2.3 Handling Protein Modifications 143 6.2.4 Handling Sample Purity 143 6.3 Bioinformatics Workflow 146 6.3.1 Spectral Preprocessing 146 6.3.2 Spectral Alignment-based Approach 146 6.3.3 Sequence Homology-based Approaches 147 6.3.4 Semi-automated and Manual de novo Sequencing 149 6.4 Sequence Validation 149 6.4.1 Mass Spectrometry-based Statistics 149 6.4.2 Intact Mass Comparison 150 6.4.3 Synthetic Peptides 150 6.5 Conclusions 150 References 151 7 Characterization of Antibody – Drug Conjugates 155; Yichin Liu 7.1 Introduction 156 7.2 Characterization of DAR Utilizing MS 157 7.2.1 The Stability of Conjugation Chemistry and the Cleavable Linker of ADC 157 7.2.2 Historical Usage of Hydrophobic Interaction Chromatography in ADC Characterization 158 7.2.3 Intact MS Detection under Denaturing Condition 159 7.2.4 Intact MS Characterization under Native Conditions 159 7.2.5 Middle-down and Bottom-up MS Approach in Mapping Drug Conjugates 161 7.3 Structural Characterization of ADC 162 7.3.1 Ion-Mobility Mass Spectrometry 162 7.3.2 Hydrogen–Deuterium Exchange Mass Spectrometry 163 7.4 Characterization of ADC Catabolism by MS 163 7.5 Conclusions 164 References 165 8 Characterization of Bispecific or Other Hybrid Molecules 169; T. Noelle Lombana and Christoph Spiess 8.1 Introduction 170 8.1.1 Bispecific Antibody Applications 170 8.2 Overview of the Various Bispecific Formats 172 8.2.1 Purification from Mixtures 175 8.2.2 Bispecific Antibodies and Alternative Scaffolds with Tethered Domains 176 8.2.3 Bispecific Molecules with Engineered Mutations 177 8.2.4 Native Bispecific IgG with Dual Binding Behavior 178 8.2.5 Bispecific Antibody Conjugates 179 8.3 Alternatives to Bispecific Antibodies: Antibody Mixtures 179 8.4 Characterization of the Bispecific Molecule 180 8.4.1 Characterization by Bioanalytical Methods 180 8.4.2 Characterization by Mass Spectrometry Methods 183 8.4.2.1 General Considerations 183 8.4.2.2 Purity Analysis of the Final Bispecific Antibody 183 8.4.2.3 Antibody Mixtures 184 8.4.2.4 Increasing Resolution 185 8.4.3 Characterization of Bispecific Antibodies by Binding Assays 185 8.4.4 Developability Assessment of the Bispecific Antibody 186 8.4.4.1 Expression 186 8.4.4.2 Physicochemical Properties 187 8.4.4.3 Chemical Modifications 187 8.4.4.4 Characterization of In Vivo Properties 188</p&gt
• Edition: 1st
• Publisher Details: Hoboken, New Jersey : John Wiley & Sons, Inc
• Publication Date: 2017
• Extent: 1 online resource
• Subjects: 615.7; Proteins -- Therapeutic use; Proteins -- Analysis
• Languages: English
• ISBNs: 9781119384403; 9781119384427
• Related ISBNs: 9781119053101
• Notes: Note: Description based on CIP data; resource not viewed.
• Access Rights: Legal Deposit; Only available on premises controlled by the deposit library and to one user at any one time; The Legal Deposit Libraries (Non-Print Works) Regulations (UK).
• Access Usage: Restricted: Printing from this resource is governed by The Legal Deposit Libraries (Non-Print Works) Regulations (UK) and UK copyright law currently in force.