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J Chromatogr A. 2018 Sep 21;1568:108-122. doi: 10.1016/j.chroma.2018.07.028. Epub 2018 Jul 6.

Evaluation of chromatofocusing as a capture method for monoclonal antibody products.

Author information

1
Department of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore County, Baltimore, MD 21250, USA.
2
Purification Process Development, Bristol-Myers Squibb, Devens, MA 01434, USA.
3
Manufacturing Sciences and Technology, Bristol-Myers Squibb, East Syracuse, NY 13057, USA.
4
Process Development Analytics, Bristol-Myers Squibb, Devens, MA 01434, USA.
5
Biologics Process Development, Bristol-Myers Squibb, Devens, MA 01434, USA.
6
Department of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore County, Baltimore, MD 21250, USA. Electronic address: dfrey1@umbc.edu.

Abstract

Chromatofocusing is investigated as an alternative to protein A chromatography for the initial capture step in a purification process for several monoclonal antibodies and antibody fusion products. For comparison, this work also investigates the use of ion-exchange chromatography with either pH or salt gradient elution as additional alternatives to protein A chromatography. The specific conditions employed for the capture step for the case of chromatofocusing were selected on a rational basis using a computer-aided design method implemented in the form of a Microsoft Excel spreadsheet. Alternative operating conditions were compared experimentally with regard to the product yield achieved as well as the removal of total host cell proteins (HCPs) and of a specific HCP major component. Results from this study indicate that both chromatofocusing and ion-exchange chromatography are useful alternatives to a protein A chromatography capture step in many practical cases. This is especially true for the case of chromatofocusing when it is possible to exploit the ability of the method to create complex gradient shapes that are self-forming inside the column and to simultaneous focus and separate proteins inside the column.

KEYWORDS:

Antibody; Chromatofocusing; Host cell proteins; Protein purification; Purification process development

PMID:
30072232
DOI:
10.1016/j.chroma.2018.07.028
[Indexed for MEDLINE]

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