Protein Analysis using Mass Spectrometry

1 - Cover [Seite 0]
2 - Title Page [Seite 5]
3 - Copyright [Seite 6]
4 - Contents [Seite 7]
5 - List of Contributors [Seite 15]
6 - Foreword [Seite 19]
7 - Preface [Seite 21]
8 - Chapter 1 Contemporary Protein Analysis by Ion Mobility Mass Spectrometry [Seite 23]
8.1 - 1.1 Introduction [Seite 23]
8.2 - 1.2 Traveling-Wave Ion Mobility Mass Spectrometry [Seite 23]
8.3 - 1.3 IM-MS and LC-IM-MS Analysis of Simple and Complex Mixtures [Seite 24]
8.3.1 - 1.3.1 Cross Section and Structure [Seite 24]
8.3.2 - 1.3.2 Separation [Seite 26]
8.3.3 - 1.3.3 Sensitivity [Seite 27]
8.4 - 1.4 Outlook [Seite 29]
8.5 - Acknowledgment [Seite 30]
8.6 - References [Seite 30]
9 - Chapter 2 High-Resolution Accurate Mass Orbitrap and Its Application in Protein Therapeutics Bioanalysis [Seite 33]
9.1 - 2.1 Introduction [Seite 33]
9.2 - 2.2 Triple Quadrupole Mass Spectrometer and Its Challenges [Seite 33]
9.3 - 2.3 High-Resolution Mass Spectrometers [Seite 34]
9.4 - 2.4 Quantitation Modes on Q Exactive Hybrid Quadrupole Orbitrap [Seite 35]
9.5 - 2.5 Protein Quantitation Approaches Using Q Exactive Hybrid Quadrupole Orbitrap [Seite 36]
9.6 - 2.6 Data Processing [Seite 38]
9.7 - 2.7 Other Factors That Impact LC-MS-based Quantitation [Seite 38]
9.7.1 - 2.7.1 Sample Extraction to Reduce Matrices [Seite 38]
9.7.2 - 2.7.2 Internal Standard [Seite 39]
9.8 - 2.8 Conclusion and Perspectives of LC-HRMS in Regulated Bioanalysis [Seite 40]
9.9 - References [Seite 40]
10 - Chapter 3 Current Methods for the Characterization of Posttranslational Modifications in Therapeutic Proteins Using Orbitrap Mass Spectrometry [Seite 43]
10.1 - 3.1 Introduction [Seite 43]
10.2 - 3.2 Characterization of PTMs Using Higher-Energy Collision Dissociation [Seite 45]
10.2.1 - 3.2.1 Oxidation [Seite 46]
10.2.2 - 3.2.2 Deamidation [Seite 46]
10.3 - 3.3 Application of Electron Transfer Dissociation to the Characterization of Labile PTMs [Seite 48]
10.3.1 - 3.3.1 Performing ETD Experiments in Orbitrap Instruments [Seite 49]
10.3.2 - 3.3.2 Structure Elucidation of Glycopeptides Using Multiple Fragmentation Mechanisms in Orbitrap Instruments [Seite 50]
10.4 - 3.4 Conclusion [Seite 53]
10.5 - Acknowledgment [Seite 54]
10.6 - References [Seite 54]
11 - Chapter 4 Macro- to Micromolecular Quantitation of Proteins and Peptides by Mass Spectrometry [Seite 57]
11.1 - 4.1 Introduction [Seite 57]
11.2 - 4.2 Key Challenges of Peptide Bioanalysis [Seite 58]
11.2.1 - 4.2.1 Key Benefits of the LC/MS/MS Peptide Quantitation Workflow [Seite 60]
11.3 - 4.3 Key Features of LC/MS/MS-Based Peptide Quantitation [Seite 60]
11.3.1 - 4.3.1 Sensitivity [Seite 61]
11.3.2 - 4.3.2 Selectivity [Seite 61]
11.3.2.1 - 4.3.2.1 MRM3 [Seite 61]
11.3.2.2 - 4.3.2.2 Differential Mobility Spectrometry (DMS) [Seite 61]
11.3.3 - 4.3.3 High?Resolution Accurate?Mass Spectrometry [Seite 61]
11.3.3.1 - 4.3.4 Software [Seite 62]
11.4 - 4.4 Advantages of the Diversity of Mass Spectrometry Systems [Seite 63]
11.5 - 4.5 Perspectives for the Future [Seite 63]
11.6 - References [Seite 64]
12 - Chapter 5 Peptide and Protein Bioanalysis Using Integrated Column-to-Source Technology for High-Flow Nanospray [Seite 67]
12.1 - 5.1 Introduction - LC-MS Has Enabled the Field of Protein Biomarker Discovery [Seite 67]
12.1.1 - 5.2 Integration of Miniaturized LC with Nanospray ESI-MS Is a Key for Success [Seite 68]
12.1.2 - 5.3 Micro- and Nano-LC Are Well Suited for Quantitative Bioanalysis [Seite 69]
12.1.3 - 5.4 Demonstrating Packed-Emitter Columns Are Suitable for Bioanalysis [Seite 71]
12.1.4 - 5.5 Future Outlook [Seite 73]
12.2 - References [Seite 74]
13 - Chapter 6 Targeting the Right Protein Isoform: Mass Spectrometry-Based Proteomic Characterization of Alternative Splice Variants [Seite 77]
13.1 - 6.1 Introduction [Seite 77]
13.2 - 6.2 Alternative Splicing and Human Diseases [Seite 77]
13.3 - 6.3 Identification of Splice Variant Proteins [Seite 78]
13.3.1 - 6.3.1 Global Profiling of Splicing Variant Proteins [Seite 78]
13.3.2 - 6.3.2 Characterization of Relative Expression of Protein Splice Variants [Seite 79]
13.3.3 - 6.3.3 Quantitation of Splice Variants by MRM-MS [Seite 84]
13.4 - 6.4 Conclusion [Seite 86]
13.5 - References [Seite 86]
14 - Chapter 7 The Application of Immunoaffinity-Based Mass Spectrometry to Characterize Protein Biomarkers and Biotherapeutics [Seite 89]
14.1 - 7.1 Introduction [Seite 89]
14.1.1 - 7.1.1 The Importance of Protein Measurement [Seite 89]
14.1.2 - 7.1.2 Ligand Binding Assays [Seite 90]
14.1.3 - 7.1.3 The Introduction of Hybrid IA-MS Methods [Seite 90]
14.2 - 7.2 Overview of IA-MS Methods [Seite 91]
14.2.1 - 7.2.1 Classification of IA-MS Methods [Seite 91]
14.2.2 - 7.2.2 Stable-Isotope-Labeled Internal Standards [Seite 93]
14.2.3 - 7.2.3 IA Capture Formats [Seite 93]
14.2.4 - 7.2.4 Liquid Chromatography [Seite 94]
14.2.5 - 7.2.5 MS Detection [Seite 96]
14.3 - 7.3 IA-MS Applications - Biomarkers [Seite 96]
14.3.1 - 7.3.1 Peptide Biomarkers [Seite 96]
14.3.2 - 7.3.2 Protein Biomarkers - Anti-Protein Capture [Seite 100]
14.3.3 - 7.3.3 Protein Biomarkers - Anti-Peptide Capture [Seite 102]
14.4 - 7.4 IA-MS Applications - Biotherapeutics [Seite 103]
14.4.1 - 7.4.1 Therapeutic Peptides [Seite 103]
14.4.2 - 7.4.2 Therapeutic Antibodies [Seite 105]
14.4.3 - 7.4.3 Antibody-Drug Conjugates [Seite 106]
14.5 - 7.5 Future Direction [Seite 106]
14.6 - References [Seite 107]
15 - Chapter 8 Semiquantification and Isotyping of Antidrug Antibodies by Immunocapture-LC/MS for Immunogenicity Assessment [Seite 113]
15.1 - 8.1 Introduction [Seite 113]
15.2 - 8.2 Multiplexing Direct Measurement of ADAs by Immunocapture-LC/MS for Immunogenicity Screening, Titering, and Isotyping [Seite 115]
15.3 - 8.3 Indirect Measurement of ADAs by Quantifying ADA Binding Components [Seite 117]
15.4 - 8.4 Use of LC-MS to Assist in Method Development of Cell-Based Neutralizing Antibody Assays [Seite 118]
15.5 - 8.5 Conclusion and Future Perspectives [Seite 119]
15.6 - References [Seite 119]
16 - Chapter 9 Mass Spectrometry-Based Assay for High-Throughput and High-Sensitivity Biomarker Verification [Seite 121]
16.1 - 9.1 Background [Seite 121]
16.2 - 9.2 Sample Processing Strategies [Seite 122]
16.3 - 9.3 Advanced Electrospray Ionization Mass Spectrometry Instrumentation [Seite 124]
16.4 - 9.4 Conclusion [Seite 127]
16.5 - References [Seite 127]
17 - Chapter 10 Monitoring Quality of Critical Reagents Used in Ligand Binding Assays with Liquid Chromatography Mass Spectrometry (LC-MS) [Seite 129]
17.1 - 10.1 Introduction [Seite 129]
17.2 - 10.2 Case Study Examples [Seite 136]
17.2.1 - 10.2.1 Case Study #1: Confirmation of Correct Reagent Construct Prior to Use in Development of an LBA Method [Seite 136]
17.2.2 - 10.2.2 Case Study #2: Monitoring the Integrity of the Reagent Cell Line Production System [Seite 138]
17.2.3 - 10.2.3 Case Study #3: Investigation of the Loss of LBA Specificity During Clinical Development [Seite 138]
17.2.3.1 - 10.2.3.1 Prestudy Investigation [Seite 138]
17.2.3.2 - 10.2.3.2 In-Study Investigation [Seite 141]
17.2.4 - 10.2.4 Case Study #4: Monitoring the Incorporation Ratio of Conjugated Critical Reagent Used in LBAs [Seite 144]
17.3 - 10.3 Discussion [Seite 144]
17.3.1 - 10.3.1 Keys to Reagent Management [Seite 144]
17.3.2 - 10.3.2 Importance of LC-MS Characterization [Seite 145]
17.3.3 - 10.3.3 The Analytical Toolbox and a "Fit-for-Purpose" Approach for Reagent Management [Seite 147]
17.4 - Acknowledgment [Seite 148]
17.5 - References [Seite 148]
18 - Chapter 11 Application of Liquid Chromatography?High Resolution Mass Spectrometry in the Quantification of Intact Proteins in Biological Fluids [Seite 151]
18.1 - 11.1 Introduction [Seite 151]
18.2 - 11.2 Workflows for Quantification of Proteins Using Full-Scan LC-HRMS [Seite 153]
18.2.1 - 11.2.1 Sample Preparation [Seite 153]
18.2.1.1 - 11.2.1.1 Solid?Phase Extraction (SPE) [Seite 153]
18.2.1.2 - 11.2.1.2 Affinity Enrichment [Seite 153]
18.2.1.3 - 11.2.1.3 Depletion of High?Abundant Proteins [Seite 153]
18.2.1.4 - 11.2.1.4 Solution Fractionation [Seite 154]
18.2.1.5 - 11.2.1.5 Protein Precipitation for PEGylated Proteins [Seite 154]
18.2.2 - 11.2.2 LC?HRMS [Seite 154]
18.2.2.1 - 11.2.2.1 HPLC [Seite 154]
18.2.2.2 - 11.2.2.2 Full?Scan HRMS Data Acquisition and Analysis [Seite 155]
18.3 - 11.3 Internal Standard Strategy [Seite 155]
18.3.1 - 11.3.1 Stable Isotope Labeled Protein [Seite 156]
18.3.2 - 11.3.2 Protein Analog [Seite 157]
18.4 - 11.4 Calibration and Quality Control (QC) Sample Strategy [Seite 157]
18.5 - 11.5 Common Issues in Quantification of Proteins Using LC-HRMS [Seite 157]
18.5.1 - 11.5.1 Stability [Seite 157]
18.5.2 - 11.5.2 Adsorption [Seite 158]
18.5.3 - 11.5.3 Specific Protein Binding [Seite 158]
18.5.4 - 11.5.4 Posttranslational Modifications (PTMs) [Seite 158]
18.6 - 11.6 Examples of LC-HRMS-Based Intact Protein Quantification [Seite 159]
18.7 - 11.7 Conclusion and Future Perspectives [Seite 160]
18.8 - Acknowledgment [Seite 162]
18.9 - References [Seite 162]
19 - Chapter 12 LC-MS/MS Bioanalytical Method Development Strategy for Therapeutic Monoclonal Antibodies in Preclinical Studies [Seite 167]
19.1 - 12.1 Introduction: LC-MS/MS Bioanalysis of Therapeutic Monoclonal Antibodies [Seite 167]
19.2 - 12.2 Highlights of Recent Method Development Strategies [Seite 168]
19.2.1 - 12.2.1 Strategy for Surrogate Peptide Selection and Optimization [Seite 168]
19.2.2 - 12.2.2 Sample Preparation [Seite 170]
19.2.2.1 - 12.2.2.1 Immunoaffinity?Based Sample Preparation [Seite 170]
19.2.2.2 - 12.2.2.2 Nonimmunoaffinity?Based Sample Preparation [Seite 173]
19.2.3 - 12.2.3 Accelerated Trypsin Digestion [Seite 174]
19.2.4 - 12.2.4 Internal Standard Selection [Seite 175]
19.2.4.1 - 12.2.4.1 SIL?Peptide IS [Seite 176]
19.2.4.2 - 12.2.4.2 Cleavable Flanking SIL?Peptide IS [Seite 176]
19.2.4.3 - 12.2.4.3 SIL?mAb IS [Seite 176]
19.3 - 12.3 Case Studies of Preclinical Applications of LC-MS/MS for Monoclonal Antibody Bioanalysis [Seite 176]
19.3.1 - 12.3.1 Case Study #1 [Seite 176]
19.3.1.1 - 12.3.1.1 Key Analytical Method Features [Seite 176]
19.3.2 - 12.3.2 Case Study #2 [Seite 177]
19.3.2.1 - 12.3.2.1 Key Analytical Method Features [Seite 177]
19.4 - 12.4 Conclusion and Future Perspectives [Seite 178]
19.5 - References [Seite 180]
20 - Chapter 13 Generic Peptide Strategies for LC-MS/MS Bioanalysis of Human Monoclonal Antibody Drugs and Drug Candidates [Seite 183]
20.1 - 13.1 Introduction [Seite 183]
20.2 - 13.2 A Universal Peptide LC-MS/MS Assay for Bioanalysis of a Diversity of Human Monoclonal Antibodies and Fc Fusion Proteins in Animal Studies1 [Seite 183]
20.2.1 - 13.2.1 Identification of a Candidate Universal Surrogate Peptide to Enable Quantification of Human mAb and Fc Fusion Protein Drug Candidates [Seite 183]
20.2.2 - 13.2.2 Application of an Exploratory Universal (Peptide 1) LC-MS/MS Assay to a Monkey Pharmacokinetic Study [Seite 184]
20.2.3 - 13.2.3 Potential Applicability of a Peptide 1 Variant to Bioanalysis of Human IgG2?Based mAbs and Fc Fusion Proteins [Seite 185]
20.2.4 - 13.2.4 Impact of Peptide 1 Asparagine Deamidation on Human mAb Quantification Can Be Mitigated [Seite 186]
20.3 - 13.3 An Improved "Dual" Universal Peptide LC-MS/MS Assay for Bioanalysis of Human mAb Drug Candidates in Animal Studies [Seite 187]
20.3.1 - 13.3.1 Identification and Evaluation of "Dual" Universal Peptide LC-MS/MS Assay Candidates [Seite 187]
20.3.2 - 13.3.2 Quantitative Evaluation and Comparison of Light and Heavy Chain Dual Universal Peptide Candidates [Seite 189]
20.3.3 - 13.3.3 Assessing the Level of Quantitative Agreement Between Peptide 1 and Peptide 2 in Assay Performance Evaluation Runs [Seite 189]
20.3.4 - 13.3.4 Deployment of the Exploratory Dual Universal Peptide Assay in Support of a Monkey Pharmacokinetic Study [Seite 190]
20.3.5 - 13.3.5 Considerations for Calibration Curve/QC Replicate Acceptance Criteria When a Dual Peptide Assay Is Employed [Seite 190]
20.3.6 - 13.3.6 Interpreting and Reporting Study Sample Concentration Data Generated with a Dual Peptide Assay [Seite 190]
20.3.7 - 13.3.7 Related Studies: Generic LC-MS/MS Assays for Human mAb Bioanalysis in Animal Studies [Seite 191]
20.4 - 13.4 Extending the Universal Peptide Assay Concept to Human mAb Bioanalysis in Human Studies [Seite 192]
20.4.1 - 13.4.1 Potential Expansion of the Universal LC-MS/MS Assay Concept into Human Studies [Seite 192]
20.4.2 - 13.4.2 Development and Evaluation of an Exploratory Universal IgG4 Clinical LC-MS/MS Assay [Seite 193]
20.4.3 - 13.4.3 Evaluation of the Impact of Anti?mAb Antibodies on Exploratory Universal IgG4 LC-MS/MS Assay Performance [Seite 195]
20.5 - 13.5 Internal Standard Options for Generic Peptide LC-MS/MS Assays [Seite 195]
20.5.1 - 13.5.1 Stable Isotopically Labeled Peptide Internal Standards [Seite 195]
20.5.2 - 13.5.2 Stable Isotopically Labeled Protein Internal Standards [Seite 196]
20.5.3 - 13.5.3 "Flanked" Stable Isotopically Labeled Peptide Internal Standards [Seite 197]
20.6 - 13.6 Sample Preparation Strategies for Generic Peptide LC-MS/MS Assays [Seite 197]
20.6.1 - 13.6.1 Direct Digestion, Pellet Digestion, and Solid?Phase Extraction [Seite 197]
20.6.2 - 13.6.2 Affinity Capture [Seite 198]
20.6.3 - 13.6.3 Additional Sample Preparation Approaches for Generic Peptide LC-MS/MS Assays [Seite 198]
20.7 - 13.7 Limitations of Generic Peptide LC-MS/MS Assays [Seite 199]
20.8 - 13.8 Conclusion [Seite 200]
20.9 - Acknowledgments [Seite 200]
20.10 - References [Seite 200]
21 - Chapter 14 Mass Spectrometry-Based Methodologies for Pharmacokinetic Characterization of Antibody Drug Conjugate Candidates During Drug Development [Seite 205]
21.1 - 14.1 Introduction [Seite 205]
21.2 - 14.2 Mechanism of Action [Seite 205]
21.2.1 - 14.2.1 Linker Chemistry [Seite 207]
21.2.2 - 14.2.2 Toxins [Seite 207]
21.2.3 - 14.2.3 ADME [Seite 207]
21.2.4 - 14.2.4 Unique Bioanalytical Challenges [Seite 207]
21.3 - 14.3 Mass Spectrometry Measurement for DAR Distribution of Circulating ADCs [Seite 208]
21.3.1 - 14.3.1 Immunocapture of ADCs from Plasma or Serum [Seite 208]
21.3.2 - 14.3.2 Deglycosylation for Captured ADCs [Seite 209]
21.3.3 - 14.3.3 Mass Spectrometry Measurement for DAR Distribution of Circulating ADCs [Seite 210]
21.4 - 14.4 Total Antibody Quantitation by Ligand Binding or LC-MS/MS [Seite 211]
21.4.1 - 14.4.1 Ligand Binding Assay [Seite 211]
21.4.2 - 14.4.2 LC-MS/MS Assay for Total Antibody Quantitation [Seite 212]
21.4.2.1 - 14.4.2.1 Predigestion Treatment [Seite 212]
21.4.2.2 - 14.4.2.2 Enzymatic Digestion [Seite 213]
21.4.2.3 - 14.4.2.3 Postdigestion Treatment [Seite 213]
21.4.2.4 - 14.4.2.4 LC-MS/MS Analysis [Seite 213]
21.4.3 - 14.4.3 Ligand Binding versus LC-MS/MS Assays [Seite 214]
21.5 - 14.5 Total Conjugated Drug Quantitation by Ligand Binding or LC-MS/MS [Seite 215]
21.5.1 - 14.5.1 Ligand Binding Assays for ADC Quantitation [Seite 215]
21.5.1.1 - 14.5.1.1 DAR?Sensitive Total Conjugated Drug Assay [Seite 215]
21.5.1.2 - 14.5.1.2 DAR?Insensitive Total Conjugated Antibody Assay [Seite 215]
21.5.2 - 14.5.2 LC-MS/MS for the Total Conjugated Drug Quantitation [Seite 216]
21.5.2.1 - 14.5.2.1 Predigestion Treatment [Seite 216]
21.5.2.2 - 14.5.2.2 Enzymatic or Chemical Digestion [Seite 216]
21.5.2.3 - 14.5.2.3 Postdigestion Treatment [Seite 217]
21.5.2.4 - 14.5.2.4 LC-MS/MS Analysis [Seite 217]
21.5.3 - 14.5.3 Ligand Binding versus LC-MS/MS [Seite 217]
21.6 - 14.6 Catabolite Quantitation by LC-MS/MS [Seite 218]
21.6.1 - 14.6.1 Sample Preparation [Seite 218]
21.6.2 - 14.6.2 LC-MS/MS Analysis [Seite 219]
21.7 - 14.7 Preclinical and Clinical Pharmacokinetic Support [Seite 219]
21.8 - 14.8 Conclusion and Future Perspectives [Seite 220]
21.9 - References [Seite 220]
22 - Chapter 15 Sample Preparation Strategies for LC-MS Bioanalysis of Proteins [Seite 225]
22.1 - 15.1 Introduction [Seite 225]
22.2 - 15.2 Sample Preparation Strategies to Improve Assay Sensitivity [Seite 227]
22.2.1 - 15.2.1 Protein Precipitation [Seite 227]
22.2.2 - 15.2.2 Solid?Phase Extraction [Seite 227]
22.2.3 - 15.2.3 Derivatization [Seite 228]
22.2.4 - 15.2.4 Depletion of High?Abundance Proteins [Seite 229]
22.2.5 - 15.2.5 Immunoaffinity Purification [Seite 230]
22.2.5.1 - 15.2.5.1 Immunocapture of a Specific Peptide [Seite 230]
22.2.5.2 - 15.2.5.2 Immunocapture of a Specific Protein [Seite 230]
22.2.5.3 - 15.2.5.3 Generic Immunocapture [Seite 232]
22.2.6 - 15.2.6 Online Sample Preparation [Seite 233]
22.3 - 15.3 Sample Preparation Strategies to Differentiate Free, Total, and ADA-Bound Proteins [Seite 235]
22.4 - 15.4 Sample Preparation Strategies to Overcome Interference from Antidrug Antibodies or Soluble Target [Seite 236]
22.5 - 15.5 Protein Digestion Strategies [Seite 236]
22.6 - 15.6. Conclusion [Seite 237]
22.7 - Acknowledgment [Seite 238]
22.8 - References [Seite 238]
23 - Chapter 16 Characterization of Protein Therapeutics by Mass Spectrometry [Seite 243]
23.1 - 16.1 Introduction [Seite 243]
23.2 - 16.2 Variants Associated with Cysteine/Disulfide Bonds in Protein Therapeutics [Seite 243]
23.2.1 - 16.2.1 Thiolation Isoforms [Seite 244]
23.2.2 - 16.2.2 Disulfide Isoforms [Seite 244]
23.2.3 - 16.2.3 Free Sulfhydryl [Seite 246]
23.2.4 - 16.2.4 Thioether/Trisulfide Bond [Seite 246]
23.2.5 - 16.2.5 Disulfide Bond in Antibody Drug Conjugates [Seite 246]
23.3 - 16.3 N-C-Terminal Variants [Seite 247]
23.4 - 16.4 Glycation [Seite 248]
23.5 - 16.5 Oxidation [Seite 248]
23.5.1 - 16.5.1 Methionine Oxidation [Seite 249]
23.5.2 - 16.5.2 Metal?Catalyzed Oxidation (MCO) [Seite 249]
23.5.3 - 16.5.3 Photooxidation [Seite 249]
23.5.4 - 16.5.4 Deamidation [Seite 250]
23.5.5 - 16.5.5 Effect of Sequence and Structure on Deamidation [Seite 250]
23.6 - 16.6 Discoloration [Seite 250]
23.7 - 16.7 Sequence Variants [Seite 252]
23.8 - 16.8 Glycosylation [Seite 254]
23.8.1 - 16.8.1 Glycoprotein Structure [Seite 254]
23.8.2 - 16.8.2 Intact Glycoprotein Analysis [Seite 257]
23.8.3 - 16.8.3 Glycopeptide Analysis [Seite 259]
23.8.4 - 16.8.4 Tandem MS of Glycopeptides [Seite 259]
23.8.5 - 16.8.5 Free Glycan Analysis [Seite 260]
23.8.6 - 16.8.6 Release of Glycans from Glycoproteins [Seite 260]
23.8.7 - 16.8.7 Detailed Sequence and Linkage Analysis of Glycans [Seite 261]
23.9 - 16.9 Conclusion [Seite 262]
23.10 - References [Seite 262]
24 - Index [Seite 273]
25 - EULA [Seite 281]