CHROMATOGRAPHIA An International Journal for Rapid Communication in Chromatography, Electrophoresis, and Associated Techniques

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Volume 53, Supplement 2001

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Scientific Editors

Technical Editors

T. A. Berger Berger Instruments Inc. 123 A Sandy Drive Newark, DE 19713, USA

K.-S. Boos Institut für Klinische Chemie

Klinikum der Universität München

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К. D. Altria, Analytical Evaluation Gp. Glaxo R&D, Ware, Herts SG12 ODP, UK

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CHROMATOGRAPHIA An International Journal for Rapid Communication in Chromatography, Electrophoresis, and Associated Techniques

Contents Supplement Vol. 53,2001

Chromatographie CE Series Series Editor: Kevin D. Altria, Analytical Evaluation Group, Glaxo R&D, Ware, Herts SG 12 ODP, UK

CE in Biotechnology: Practical Applications for Protein and Peptide Analyses Edited by Anthony B. Chen, Genentech Inc. South San Francisco, CA, USA

Wassim Nashabeh Genentech Inc. South San Francisco, CA, USA

Timothy Wehr

Bay Bioanalytical Laboratory, Richmond, CA, USA

Preface S -5

1 Capillary Electrophoresis in the Development of Recombinant Protein Biopharmaceuticals S -7 A. B. Chen / E. Canova-Davis

1.1 Summary S -7 1.2 Introduction S-7 1.3 Capillary Electrophoresis Reviews S-8 1.4 Reviews of CE Used for the Analysis

of Recombinant Proteins S-ll 1.5 Recent CE Literature S-13 1.6 Quality Control and Validation S-15 1.7 Developments in CE-MS S-15 1.8 References S-16 2 Selection of Buffers in Capillary Zone Electrophoresis:

Application to Peptide and Protein Analysis S-18 G. M. Janini / H. J. Issaq

2.1 Introduction S-18 2.2 Buffer Parameters S-18 2.3 Buffers Used for the Separation of Peptides

and Proteins S -23 2.4 Acknowledgements S -25 2.5 References S -25

3 Reverse-Phase High Performance Liquid Chromatography and Capillary Zone Electrophoresis Separation of Protegrin Analogs S -27 J. Chen / J. Fausnaugh-Pollitt

3.1 Abstract S-27 3.2 Introduction S-27 3.3 Experimental S-28 3.4 Results and Discussion S -29 3.5 References S -33

4 Use of Capillary Electrophoresis to Determine the Dilute Protein Concentration in Formulations Containing Interfering Excipients S-34 S. S. Park / A. Cate / B. S. Chang

4.1 Abstract S-34 4.2 Introduction S-34 4.3 Material and Methods S-35 4.4 Result and Discussion S-35 4.5 Conclusion S-38 4.6 Acknowledgements S-38 4.7 References S-38

5 Effects of the Solution Environment on the Resolution of Recombinant Human Deoxyribonuclease Variants in Capillary Zone Electrophoresis S -39 С P. Quan / E. Canova-Davis /A .B . Chen Abstract S-39 Introduction S -39 Materials S-39 Methods S-40 Results and Discussion S-40 Conclusion S-44 Acknowledgements S-44 References S-44

Recent Advances in Capillary Isoelectric Focusing T. Wehr / R. Rodriguez-Diaz / M. Zhu S -45 Introduction S-45 Theory and Practice of cIEF S-45 cIEF in Microchannels S - 47 ImagingcIEF S-48 cIEF-Mass-Spectrometry S-49 Capillary Isoelectric Focusing of Monoclonal Antibodies S -50 Capillary Isoelectric Focusing of Hemoglobin and Hemoglobin Variants S-54 Capillary Isoelectric Focusing of Erythropoietin.... S -57 Novel Applications of cIEF S-57 References S -58

7 Separation of Enbrel® (rhuTNFR: Fc) Isoforms by Capillary Isoelectric Focusing S -59 С Jochheim / S. Novick / A. Balland / J. Mahan-Boyce / W.-C. Wang / A. Goetze / W. Gombotz

7.1 Abstract S-59 7.2 Introduction S-59 7.3 Experimental S -60

5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8

6.1 6.2 6.3 6.4 6.5

6.7

6.8 6.9 6.10

S-2 Chromatographia Supplement Vol. 53,2001 Contents

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7.4 Results and Discussion S -61 7.5 Acknowledgements S -64 7.6 References S -65

8 , Optimization, Validation, and Use of Capillary Gel Electrophoresis for Quality Control Testing of Synagis®, a Monoclonal Antibody S -66 M. A. Schenerman / S. H. Bowen

8.1 Abstract S-66 8.2 Introduction S -66 8.3 Methods S-67 8.4 Regulatory Guidance S-67 8.5 Optimization Strategy S -68 8.6 Validation S-68 8.7 Validation Parameters for CGE S-68 8.8 Acknowledgements S -72 8.9 References S -72

9 Analysis of Protein Therapeutics by Capillary Electrophoresis S -75 S.Ma/W.Nashabeh

9.1 Introduction S-75 9.2 Capillary Electrophoresis - Sodium Dodecylsulfate

(CE-SDS) S-75

9.3 Capillary Isoelectric Focusing (cIEF) S -81 9.4 Capillary Zone Electrophoresis (CZE) S-83 9.5 Carbohydrate Analysis S-84 9.6 Methods S-86 9.7 Acknowledgements S -89 9.8 References S-89

10 Sensitive and High Resolution CE/MS/MS for Protein Identification in Complex Mixtures S -90 W.Tong/J. R.Yates

10.1 Introduction S-90 10.2 Approaches to Improve the Sensitivity

of CE/MS/MS for Protein Identification S-90 10.3 Approaches to Improve the Resolution

of CE/MS/MS for Protein Identification S -94 10.4 Other Approaches S-96 10.5 Conclusions S-97 10.6 References S-98

Subject Index S-100

Abbreviations S -102

Contents Chromatographia Supplement Vol. 53, 2001 S-3