Salt tolerant membrane adsorbers for robust impurity clearance
| Autor: | Steven M C O Minnesot Heilmann, Mark R. Etzel, Kurt Brorson, William Riordan, Kannan Seshadri |
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| Rok vydání: | 2009 |
| Předmět: |
medicine.drug_class
Inorganic chemistry Salt (chemistry) CHO Cells Monoclonal antibody Ligands Adsorption Cricetulus Cricetinae medicine Animals Bacteriophages Anion Exchange Resins chemistry.chemical_classification Chromatography Ion exchange Hydrogen bond Ligand Antibodies Monoclonal Proteins Membranes Artificial Serum Albumin Bovine Hydrogen-Ion Concentration Chromatography Ion Exchange Membrane chemistry Amine gas treating Salts Drug Contamination Biotechnology |
| Zdroj: | Biotechnology progress. 25(6) |
| ISSN: | 1520-6033 |
| Popis: | Clearance of impurities such as viruses, host cell protein (HCP), and DNA is a critical purification design consideration for manufacture of monoclonal antibody therapeutics. Anion exchange chromatography has frequently been utilized to accomplish this goal; however, anion exchange adsorbents based on the traditional quaternary amine (Q) ligand are sensitive to salt concentration, leading to reduced clearance levels of impurities at moderate salt concentrations (50-150 mM). In this report, membrane adsorbers incorporating four alternative salt tolerant anion exchange ligands were examined for impurity clearance: agmatine, tris-2-aminoethyl amine, polyhexamethylene biguanide (PHMB), and polyethyleneimine. Each of these ligands provided greater than 5 log reduction value (LRV) for viral clearance of phage phiX174 (pI approximately 6.7) at pH 7.5 and phage PR772 (pI approximately 4) at pH 4.2 in the presence of salt. Under these same conditions, the commercial Q membrane adsorber provided no clearance (zero LRV). Clearance of host-cell protein at pH 7.5 was the most challenging test case; only PHMB maintained 1.5 LRV in 150 mM salt. The salt tolerance of PHMB was attributed to its large positive net charge through the presence of multiple biguanide groups that participated in electrostatic and hydrogen bonding interactions with the impurity molecules. On the basis of the results of this study, membrane adsorbers that incorporate salt tolerant anion exchange ligands provide a robust approach to impurity clearance during the purification of monoclonal antibodies. |
| Databáze: | OpenAIRE |
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