Optimization of protein G chromatography for bio pharmaceutical monoclonal antibodies
| Autor: | Eberhard Bill, Britt-Marie Karlsson, Ulrike Lutz, Marianne Sparrman, Hermann Allgaier |
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| Rok vydání: | 1995 |
| Předmět: |
medicine.drug_class
Fast flow Ligands Monoclonal antibody Chromatography Affinity Sepharose Matrix (chemical analysis) Acetic acid chemistry.chemical_compound Bacterial Proteins Structural Biology medicine Humans Molecular Biology Chromatography biology Elution Regeneration (biology) Antibodies Monoclonal Streptococcus Molecular Weight chemistry Immunoglobulin G biology.protein Electrophoresis Polyacrylamide Gel Indicators and Reagents Protein G |
| Zdroj: | Journal of Molecular Recognition. 8:90-94 |
| ISSN: | 1099-1352 0952-3499 |
| DOI: | 10.1002/jmr.300080116 |
| Popis: | Compared to more stable chromatographic media the use of affinity media with biological ligands such as Protein G Sepharose® 4 Fast Flow poses special challenges regarding regeneration and sanitization. This is especially critical for the purification of pharmaceutical proteins, where complete regeneration of the column between runs is of paramount importance. Here, the problems encountered during process development and up scaling of regeneration methods for a protein G Sepharose Fast Flow column intended for the large-scale purification of pharmaceutical monoclonal anitbiodies are reported. The initially chosen alkaline regeneration buffer led to an increase in the affinity of Protein G towards antibodies which made elution increasingly difficult. A combination of urea and acetic acid was selected to ensure efficient cleaning of the matrix without affecting ligand properties. Validation experiments were done to demonstrate the functional integrity of the matrix after repeated cycles of use and regeneration, as well as the efficiency of the cleaning process. |
| Databáze: | OpenAIRE |
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