![]() ![]() 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. Capto Phenyl (High sub) and Capto Butyl are hydrophobic interaction chromatography (HIC) media used in the capture and intermediate stages of protein purification Prepacked with Capto Phenyl (high sub) Excellent for method optimization and parameter screening due to the 10 cm bed height Easily. Furthermore, the impact of salt removal from the HIC load on protein-resin binding was examined providing opportunities to minimize processing time. 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. In some aspects, a hydrophobic strength of the HIC column of the. Capto Phenyl High Sub 25 ml is a hydrophobic interaction chromatography (HIC) medium used in the capture and intermediate stages of protein purification. The influences of gradient versus step elution as well as superficial linear velocity on product quality were evaluated. In some aspects, a resin of the HIC column of the method is phenyl or capto phenyl resin. Results and discussion Hydrophobic interaction chromatography is routinely used for the purification and manufacture of bio-pharmaceuticals. Absorbance at 280 nm was used to determine peak collection in both cases. Capto Phenyl (high sub) is ideal for initial and intermediate step purifications requiring a chromatography resin with medium to high hydrophobicity. 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. Capto Phenyl ImpRes resin was used to purify Mab 2 under similar conditions but performed at 200 cm/h. 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. Hydrophobic interaction chromatography (HIC) is routinely used in the purification of biopharmaceuticals such as antibodies. ![]()
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