Protein A chromatography is the industry gold-standard for monoclonal antibody and Fc-fusion protein capture. The high cost of Protein A however makes resin lifetime and resin reuse an important factor for process economics.  Resin lifetime studies usually examine the effect of resin re-use on binding capacity, yield, and product quality without answering the fundamental question of what is causing the decrease in performance.  A two part mechanistic study was conducted to decouple the effect of the two possible factors (ligand hydrolysis and/or degradation vs. resin fouling) on column performance over lifetime of the most commonly used base-stable Protein A resins (MabSelect SuRe and MabSelect SuRe LX).  We examined the change in binding capacity as a function of sodium hydroxide concentration (rate of hydrolysis), temperature, and stabilizing additives.  Additionally, product cycling and resin extraction studies were conducted to determine cleaning effectiveness (resin fouling) of various cleaning strategies.  Our results demonstrate that hydrolysis studies alone are not a good predictor of actual column lifetime due to resin fouling experienced during actual column cycling studies.  Sodium hydroxide based cleaning solutions were found to be most effective at preventing resin fouling while cold temperature and the use of stabilizing additives in conjunction with sodium hydroxide was found to be beneficial in minimizing the rate of Protein A ligand hydrolysis.  Overall, an effective and robust cleaning strategy was identified that could double the amount of achievable cycles at manufacturing scale.

Maximizing the functional lifetime of Protein A resins

253rd National Meeting (2017)

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