A mechanistic study of Protein A chromatography resin lifetime

Jiang, Canping; Liu, Jing; Rubacha, Michael; Shukla, Abhinav A.

doi:10.1016/j.chroma.2009.06.013

Cited by 76 publications

(87 citation statements)

References 18 publications

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“…It is important to mention that H for any given chromatographic stationary phase is product specific and depends on a variety of factors including (a) resin type, (b) concentration and nature of the feed, (c) regeneration procedures, and (d) storage conditions, among others. 37 And thus, it is empirically determined from resin life time studies. Such studies could be guided using the information presented in Figures 4A,B, as from these figures a target H can be defined based on a desired P C .…”

Section: Sensitivity Analysismentioning

confidence: 99%

Techno‐economic evaluation of an inclusion body solubilization and recombinant protein refolding process

Freydell

Wielen

Eppink

et al. 2011

Biotechnology Progress

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Expression of recombinant proteins in Escherichia coli is normally accompanied by the formation of inclusion bodies (IBs). To obtain the protein product in an active (native) soluble form, the IBs must be first solubilized, and thereafter, the soluble, often denatured and reduced protein must be refolded. Several technically feasible alternatives to conduct IBs solubilization and on-column refolding have been proposed in recent years. However, rarely these on-column refolding alternatives have been evaluated from an economical point of view, questioning the feasibility of their implementation at a preparative scale. The presented study assesses the economic performance of four distinct process alternatives that include pH induced IBs solubilization and protein refolding (pH_IndSR); IBs solubilization using urea, dithiothreitol (DTT), and alkaline pH followed by batch size-exclusion protein refolding; inclusion bodies (IBs) solubilization using urea, DTT, and alkaline pH followed by simulated moving bed (SMB) size-exclusion protein refolding, and IBs solubilization using urea, DTT and alkaline pH followed by batch dilution protein refolding. The economic performance was judged on the basis of the direct fixed capital, and the production cost per unit of product (P(C)). This work shows that (1) pH_IndSR system is a relatively economical process, because of the low IBs solubilization cost; (2) substituting β-mercaptoethanol for dithiothreithol is an attractive alternative, as it significantly decreases the product cost contribution from the IBs solubilization; and (3) protein refolding by size-exclusion chromatography becomes economically attractive by changing the mode of operation of the chromatographic reactor from batch to continuous using SMB technology.

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Section: Sensitivity Analysismentioning

confidence: 99%

Techno‐economic evaluation of an inclusion body solubilization and recombinant protein refolding process

Freydell

Wielen

Eppink

et al. 2011

Biotechnology Progress

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“…A decay of resin lifetime can be contributed by a few factors: ligand degradation, crude loading material [14,15] and ligand leakage [16] etc. Other studies have identified novel additives such as sucrose, xylitol, and ethylene glycol.…”

Section: Introductionmentioning

confidence: 99%

Effect of cleaning agents and additives on Protein A ligand degradation and chromatography performance

Yang

Harding

Иванов

et al. 2015

Journal of Chromatography A

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“…There have been various case studies reported where PAT has been successfully implemented for various downstream unit operations like protein refolding, filtration, inline mixing, protein pegylation, viral inactivation, and process chromatography . Currently, various techniques such as UV spectroscopy, HPLC, and mass spectroscopy are available to monitor column effluent and provide useful information about product purity and impurity clearance . However, these are all indirect measures of performance monitoring.…”

Section: Introductionmentioning

confidence: 99%

Implementation of a fluorescence based PAT control for fouling of protein A chromatography resin

Pathak

Rathore

2017

J of Chemical Tech & Biotech

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BACKGROUND Protein A chromatography fouling is accompanied by two major events, one is the loss of protein A ligands and other is fouling due to non‐specific, irreversible interactions of foulants with resin particles. This paper presents implementation of process analytical technology based control for fouling of protein A chromatography resin using a novel, fluorescence based approach. This approach enables direct, in situ measurement of protein A ligand density as well as monitoring of resin fouling during resin reuse. RESULTS A novel, fluorescence based process analytical technology (PAT) tool has been designed and used for screening a variety of cleaning protocols. A two‐step cleaning protocol was created using this methodology. The above mentioned fluorescence based approach was successfully used to monitor fouling and to take a decision on when to initiate cleaning. This resulted in effective maintenance of dynamic binding capacity and step yield at 50 cycles (DBC: 97% of the original value vs 65% with conventional protocols and yield: 93% of the original value vs 73% with conventional protocols) and at 200 cycles (> 90% of the original value). CONCLUSIONS The proposed fluorescence based approach has been effectively used for monitoring and control of resin fouling upon reuse, thereby resulting in a substantial increase in resin lifetime. © 2017 Society of Chemical Industry

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A mechanistic study of Protein A chromatography resin lifetime

Cited by 76 publications

References 18 publications

Techno‐economic evaluation of an inclusion body solubilization and recombinant protein refolding process

Techno‐economic evaluation of an inclusion body solubilization and recombinant protein refolding process

Effect of cleaning agents and additives on Protein A ligand degradation and chromatography performance

Implementation of a fluorescence based PAT control for fouling of protein A chromatography resin

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