NDLI: Optimization of CE-SDS Method for Antibody Separation Based on Multi-Users Experimental Practices

Content Provider	Springer Nature Link
Author	Han, M. Phan, D. Nightlinger, N. Taylor, L. Jankhah, S. Woodruff, B. Yates, Z. Freeman, S. Guo, A. Balland, A. Pettit, D.
Copyright Year	2006
Abstract	Capillary Electrophoresis-Sodium Dodecyl Sulfate (CE-SDS) method with UV detection was developed and satisfactorily used for determination of purity and manufacturing consistency of a monoclonal antibody (MAb) at Amgen Inc. (Seattle, WA). When this method was applied to some other MAbs, several problems with method robustness became apparent. These issues resulted in abnormal Electropherogram (e-gram) profiles potentially linked to various parameters specific molecules analyzed, sample formulation buffer composition, CE-SDS gel matrix type, and operators. A multi-users interest group (called CE Users Forum) was formed to systematically investigate and understand these issues. The CE Users Forum first identified the issues which needed resolution, defined group experiments to better understand the problem and to test potential solutions, and together defined a generic (platform) CE-SDS method for MAbs. Two CE instruments, Agilent HP3DCE and Beckman PA 800, two CE-SDS gel matrices, BioRad and Beckman gels, as well as different types of MAbs in various buffers were used in this investigation. We present here a platform CE-SDS method for purity determination of MAbs. Method optimization and trouble-shooting procedures by the CE Users Forum played a key role in delivering a robust analytical method for characterization of antibodies by improving instrumental and experimental parameters such as instrument variability, instrument operating parameters, operator training, and reagent stability. The optimized CE-SDS method is used during process development and has been transferred to the quality control (QC) lab as a purity assay for lot release testing of therapeutic antibodies. Any trained analyst can successfully perform this method. A group such as the CE Users Forum is a good way to integrate best practices and solve technical issues in a cooperative environment.
Starting Page	1
Ending Page	8
Page Count	8
File Format	PDF
ISSN	00095893
Journal	Chromatographia
Volume Number	64
Issue Number	5-6
e-ISSN	16121112
Language	English
Publisher	Vieweg Verlag
Publisher Date	2006-06-28
Publisher Place	Wiesbaden
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Capillary electrophoresis-sodium dodecyl sulfate Capillary gel electrophoresis Monoclonal antibody separation Troubleshooting Optimization Measurement Science, Instrumentation Plant Sciences Biochemistry Pharmacy Organic Chemistry Analytical Chemistry
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Analytical Chemistry Biochemistry Clinical Biochemistry

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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