A safe, effective, and facility compatible cleaning in place procedure for affinity resin in large-scale monoclonal antibody purification

Wang, Lu; Dembecki, J.; Jaffe, Neil; O’Mara, Brian W.; Cai, Hui; Sparks, Colleen; Zhang, Jian; Laino, Sarah; Russell, Reb J.; Wang, Michelle

doi:10.1016/j.chroma.2013.07.096

Cited by 18 publications

(15 citation statements)

References 16 publications

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“…MabSelect SuRe LX has the same functional alkali stable ligand but at much higher density on the bead matrix allowing for higher binding capacities . This permits for the use of more stringent cleaning agents, such as sodium hydroxide, which were once off limits to the Protein A step . Sodium hydroxide is both cost effective and offers no disposal restrictions at large scale manufacturing.…”

Section: Introductionmentioning

confidence: 99%

“…23,24 This permits for the use of more stringent cleaning agents, such as sodium hydroxide, which were once off limits to the Protein A step. 25,26 Sodium hydroxide is both cost effective and offers no disposal restrictions at large scale manufacturing. It has been historically used as the cleaning solution for other chromatographic steps, such as ion exchange and hydrophobic interaction chromatography 27,28 but was minimally used for Protein A due to its detrimental effect on the ligand.…”

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confidence: 99%

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Maximizing the functional lifetime of Protein A resins

Zhang

Siva

Caple

et al. 2017

Biotechnology Progress

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Protein A chromatography is currently the industry gold-standard for monoclonal antibody and Fc-fusion protein purification. The high cost of Protein A, however, makes resin lifetime and resin reuse an important factor for process economics. Typical resin lifetime studies performed in the industry 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 in an attempt to decouple the effect of the two possible factors (resin hydrolysis and/or degradation vs. resin fouling) on column performance over lifetime of the most commonly used alkali-stable Protein A resins (MabSelect SuRe and MabSelect SuRe LX). The change in binding capacity as a function of sodium hydroxide concentration (rate of hydrolysis), temperature, and stabilizing additives was examined. Additionally, resin extraction studies and product cycling studies were conducted to determine cleaning effectiveness (resin fouling) of various cleaning strategies. Sodium hydroxide-based cleaning solutions were shown to be more effective at preventing resin fouling. Conversely, cold temperature and the use of stabilizing additives in conjunction with sodium hydroxide were found to be beneficial in minimizing the rate of Protein A ligand hydrolysis. An effective and robust cleaning strategy is presented here to maximize resin lifetime and thereby the number of column cycles for future manufacturing processes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:708-715, 2017.

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Section: Introductionmentioning

confidence: 99%

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confidence: 99%

Maximizing the functional lifetime of Protein A resins

Zhang

Siva

Caple

et al. 2017

Biotechnology Progress

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“…A reducing solution followed by a chaotropic solution also proved an effective CIP strategy 28 33 . This alkaline treatment extends resin lifespan, but it also appears to decrease the binding capacity 26 due to either Protein A leaching 4 34 35 or ligand denaturation 36 . Under alkaline conditions, asparagine and glutamine residues in Protein A are susceptible to deamidation which also decreases binding capacity 37 38 .…”

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confidence: 99%

In-column ATR-FTIR spectroscopy to monitor affinity chromatography purification of monoclonal antibodies

Boulet-Audet

Kazarian

Byrne

2016

Sci Rep

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In recent years many monoclonal antibodies (mAb) have entered the biotherapeutics market, offering new treatments for chronic and life-threatening diseases. Protein A resin captures monoclonal antibody (mAb) effectively, but the binding capacity decays over repeated purification cycles. On an industrial scale, replacing fouled Protein A affinity chromatography resin accounts for a large proportion of the raw material cost. Cleaning-in-place (CIP) procedures were developed to extend Protein A resin lifespan, but chromatograms cannot reliably quantify any remaining contaminants over repeated cycles. To study resin fouling in situ, we coupled affinity chromatography and Fourier transform infrared (FTIR) spectroscopy for the first time, by embedding an attenuated total reflection (ATR) sensor inside a micro-scale column while measuring the UV 280 nm and conductivity. Our approach quantified the in-column protein concentration in the resin bed and determined protein conformation. Our results show that Protein A ligand leached during CIP. We also found that host cell proteins bound to the Protein A resin even more strongly than mAbs and that typical CIP conditions do not remove all fouling contaminants. The insights derived from in-column ATR-FTIR spectroscopic monitoring could contribute to mAb purification quality assurance as well as guide the development of more effective CIP conditions to optimise resin lifespan.

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“…Citrate showed stronger protection than sulfate because citrate is more kosmotropic than sulfate. Wang et al showed that sulfate demonstrated more protective effect than chloride in a similar experiment as predicted by Hofmeister series [17].…”

Section: Column Performance Studies-recoverymentioning

confidence: 78%

“…Other studies have identified novel additives such as sucrose, xylitol, and ethylene glycol. In another study [17] additives such as sodium sulfate and sodium chloride have been evaluated with some improvement in protection of Protein A during cleaning. While additives such as sodium chloride and sodium sulfate might provide some protection, in a manufacturing setting it is desirable to identify non-corrosive chemicals.…”

Section: Introductionmentioning

confidence: 98%