CE in Biotechnology: Practical Applications for Protein and Peptide Analyses

Anthony B. Chen, Ph.D. has worked for over fifteen years at Genentech Inc. South San Francisco, CA developing analytical assays for recombinant protein pharmaceuticals. He introduced the use of novel analytical applications, including CE, for routine protein analyses in the Quality Control department.Wassim Nashabeh, Ph.D. has worked with CE for over ten years, including pre- and postdoctoral studies and CE for commercial applications. He is Associate Director, Quality Control, Genentech Inc. where he has validated several CE methods used in the lot release of marketed protein therapeutics.Timothy Wehr, Ph.D. developed CE instrumentation and application kits as manager of the CE chemistry R&D group at Bio-Rad laboratories, Richmond, CA and has developed CE methods for biopharmaceuticals at Bay Bioanalytical Laboratory, Richmond CA.

• 1 Capillary Electrophoresis in the Development of Recombinant Protein Biopharmaceuticals.- 1.1 Summary.- 1.2 Introduction.- 1.3 Capillary Electrophoresis Reviews.- 1.4 Reviews of CE Used for the Analysis of Recombinant Proteins.- 1.5 Recent CE Literature.- 1.6 Quality Control and Validation.- 1.7 Developments in CE-MS.- 1.8 References.- 2 Selection of Buffers in Capillary Zone Electrophoresis: Application to Peptide and Protein Analysis.- 2.1 Introduction.- 2.2 Buffer Parameters.- 2.3 Buffers Used for the Separation of Peptides and Proteins.- 2.4 Acknowledgements.- 2.5 References.- 3 Reverse-Phase High Performance Liquid Chromatography and Capillary Zone Electrophoresis Separation of Protegrin Analogs.- 3.1 Abstract.- 3.2 Introduction.- 3.3 Experimental.- 3.4 Results and Discussion.- 3.5 References.- 4 Use of Capillary Electrophoresis to Determine the Dilute Protein Concentration in Formulations Containing Interfering Excipients.- 4.1 Abstract.- 4.2 Introduction.- 4.3 Material and Methods.- 4.4 Result and Discussion.- 4.5 Conclusion.- 4.6 Acknowledgements.- 4.7 References.- 5 Effects of the Solution Environment on the Resolution of Recombinant Human Deoxyribonuclease Variants in Capillary Zone Electrophoresis.- 5.1 Abstract.- 5.2 Introduction.- 5.3 Materials.- 5.4 Methods.- 5.5 Results and Discussion.- 5.6 Conclusion.- 5.7 Acknowledgements.- 5.8 References.- 6 Recent Advances in Capillary Isoelectric Focusing.- 6.1 Introduction.- 6.2 Theory and Practice of cIEF.- 6.3 cIEF in Microchannels.- 6.4 Imaging cIEF.- 6.5 cIEF-Mass-Spectrometry.- 6.6 Capillary Isoelectric Focusing of Monoclonal Antibodies.- 6.7 Capillary Isoelectric Focusing of Hemoglobin and Hemoglobin Variants.- 6.8 Capillary Isoelectric Focusing of Erythropoietin.- 6.9 Novel Applications of cIEF.- 6.10 References.- 7 Separation of Enbrel® (rhuTNFR: Fc) Isoforms by Capillary Isoelectric Focusing.- 7.1 Abstract.- 7.2 Introduction.- 7.3 Experimental.- 7.4 Results and Discussion.- 7.5 Acknowledgements.- 7.6 References.- 8 Optimization, Validation, and Use of Capillary Gel Electrophoresis for Quality Control Testing of Synagis®, a Monoclonal Antibody.- 8.1 Abstract.- 8.2 Introduction.- 8.3 Methods.- 8.4 Regulatory Guidance.- 8.5 Optimization Strategy.- 8.6 Validation.- 8.7 Validation Parameters for CGE.- 8.8 Acknowledgements.- 8.9 References.- 9 Analysis of Protein Therapeutics by Capillary Electrophoresis.- 9.1 Introduction.- 9.2 Capillary Electrophoresis — Sodium Dodecylsulfate (CE-SDS).- 9.3 Capillary Isoelectric Focusing (cIEF).- 9.4 Capillary Zone Electrophoresis (CZE).- 9.5 Carbohydrate Analysis.- 9.6 Methods.- 9.7 Acknowledgements.- 9.8 References.- 10 Sensitive and High Resolution CE/MS/MS for Protein Identification in Complex Mixtures.- 10.1 Introduction.- 10.2 Approaches to Improve the Sensitivity of CE/MS/MS for Protein Identification.- 10.3 Approaches to Improve the Resolution of CE/MS/MS for Protein Identification.- 10.4 Other Approaches.- 10.5 Conclusions.- 10.6 References.- Abbreviations.