Use of mobile phase additives for the elution of bispecific and monoclonal antibodies from phenyl based hydrophobic interaction chromatography resins
| Autor: | Warren R. Emery, Gerard M. Kelly, Troii Hall |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
Arginine medicine.drug_class Clinical Biochemistry Salt (chemistry) CHO Cells Monoclonal antibody 01 natural sciences Biochemistry Analytical Chemistry Cricetulus 010608 biotechnology Phase (matter) Cricetinae medicine Animals chemistry.chemical_classification Chromatography Elution Hydrophilic interaction chromatography Sepharose 010401 analytical chemistry Antibodies Monoclonal Proteins Cell Biology General Medicine 0104 chemical sciences chemistry Hexylene glycol Early phase Hydrophobic and Hydrophilic Interactions Chromatography Liquid |
| Zdroj: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 1096 |
| ISSN: | 1873-376X |
| Popis: | Hydrophobic interaction chromatography (HIC) is routinely used in the purification of biopharmaceuticals such as antibodies. However, hydrophobic proteins can be difficult to elute resulting in low recovery of product thereby complicating early phase process development and potentially excluding the use of HIC resins for their manufacture. Mobile phase additives such as hexylene glycol and arginine facilitate protein elution from resins including HIC; therefore, these additives were evaluated toward the recovery and purification of bispecific and monoclonal antibodies from Phenyl Sepharose HP and Capto Phenyl ImpRes resins. The influences of gradient versus step elution as well as superficial linear velocity on product quality were evaluated. Improved protein recovery and reduction of both soluble product aggregate and host cell protein were observed for the tested antibodies with both hexylene glycol and arginine. Furthermore, the impact of salt removal from the HIC load on protein-resin binding was examined providing opportunities to minimize processing time. This method was successfully scaled using a Phenyl Sepharose HP (5 cm i.d. × 20.0 cm) and Capto Phenyl ImpRes (3.2 cm i.d. × 21.4 cm) column demonstrating potential for manufacturing purposes. |
| Databáze: | OpenAIRE |
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