Riske, F., Smith, M., Menon, M., Goetschalck, S., Goidsenhoven, I. V., Krul, A., Pimpaneau, V., Renaers, I., Van Tichelt, N., Van Houdt, K., Hayes, M., Lawrence, C., Bigelow, R., Schroeder, J. B. (2007) A potential generic downstream process using Cibracon Blue resin at very high loading capacity produces a highly purified monoclonal antibody preparation from cell culture harvest. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 848 (1). pp. 108-115. ISSN 15700232
Abstract
The use of a dye-ligand chromatography for the purification of monoclonal antibody (MAb) from cell culture and other feed streams has been largely overlooked in large scale production. Cibracon Blue dye (CB), a polycyclic anionic ligand, interacts with protein through a specific interaction between the dye, acting as a mimic of NAD+ and NADP+, or through non-specific electrostatic, hydrophobic, and other forces. In this paper, a CB resin was used to effectively and efficiently separate an IgG4 MAb from host and process impurities following the capture of the MAb on a Protein-A (PA) column. The CB unit operation, challenged at ≤180 g MAb/L of resin with the PA eluate, reduced BSA (1-2 log), host cell protein (HCP; 2-3 log), MAb oligomer (31-85%), fragment (from ∼0.8% to <0.1%), and other undesired MAb species. Purity, as measured by non-reducing (NR) SDS-PAGE, was improved 33-85%, to 92-99.5% overall (>99% by reducing SDS-PAGE). A facile three column scalable production scheme, employing CB as the second column in the process was used to generate highly purified MAb from cell culture harvest derived from two media of very different compositions. Free CB dye was ≤1 ng/mg in MAb preparations purified through the three column process and then concentrated and buffer exchanged into the appropriate buffer using tangential flow filtration (TFF). © 2006 Elsevier B.V. All rights reserved.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Cibracon Blue resin Generic downstream process Monoclonal antibody Purification of cell culture harvest Feed streams Generic downstreams Polycyclic anionic ligands Tangential flow filtration (TFF) Cell culture Chromatography Dyes Hydrophobicity Monoclonal antibodies Purification Resins cell protein cibracon blue dye immunoglobulin g4 monoclonal antibody oligomer protein A resin unclassified drug article downstream processing drug manufacture drug purification drug purity drug synthesis filtration host cell polyacrylamide gel electrophoresis priority journal Animals Antibodies, Monoclonal Cell Line, Tumor Culture Media, Conditioned Electrophoresis, Polyacrylamide Gel Immunoglobulin G Resins, Synthetic |
| Subjects: | biotechnology biotechnology > chromatography biotechnology > chromatography > pseudo-affinity chromatography > dye ligand chromatography bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > monoclonal antibody biotechnology > chromatography > pseudo-affinity chromatography |
| CSHL Authors: | |
| Depositing User: | CSHL Librarian |
| Date: | 2007 |
| Date Deposited: | 06 Dec 2011 22:27 |
| Last Modified: | 05 Dec 2019 20:07 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/23249 |
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