A tool for increasing the lifetime of chromatography resins

Grönberg, Anna; Eriksson, Malin; Ersoy, Maria; Johansson, Hans J.

doi:10.4161/mabs.3.2.14874

Cited by 36 publications

(32 citation statements)

References 2 publications

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“…Protein A resins are used extensively in the purification of monoclonal antibodies (mAbs) at manufacturing scale (Boschetti and Jungbauer, ; Hober et al, ; Kelley, ; Shukla et al, ). Due to their high cost, the ability to reuse these resins is critical for economic reasons (Grönberg et al, ; Hahn et al, ; Jiang et al, ). Sodium hydroxide is often the cleaning agent of choice for many Protein A resins as it has the ability to hydrolyze proteinaceous residues and simultaneously sanitize the resin (Burgoyne et al, ; Girot et al, ; Hober et al, ).…”

Section: Introductionmentioning

confidence: 99%

Nature of foulants and fouling mechanism in the Protein A MabSelect resin cycled in a monoclonal antibody purification process

Zhang

Daniels

Salm

et al. 2015

Biotech & Bioengineering

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The composition and origin of foulants and their spatial distribution within the particles of the Protein A MabSelect resin cycled in a mAb purification process are determined using electron and confocal microscopy techniques with gold and fluorescently labeled protein probes that associate with the foulants. The results show that the foulants are primarily related to the mAb product, are heterogeneously dispersed both on the outer surface and in the interior of the resin beads, and accumulate only when loading the conditioned CHO cell culture supernatant. Insignificant accumulation is seen if the process is run with purified mAb or with the null cell culture supernatant. When bound to the Protein A ligand, the mAb responsible for the observed fouling behavior is shown to associate with BSA and α-lactalbumin. This property is exploited using labeled versions of these lipophilic proteins to assess the effectiveness of improved resin cleaning processes and to elucidate the fouling mechanism. Resin fouling for this mAb appears to be consistent with the occurrence of conformational changes that occur upon binding, which, in turn, facilitate association of lipophilic proteins with the mAb. Upon desorption at low pH, these destabilized mAb complexes are deposited on and within the resin growing with each cycle and eventually leading to significant degradation of process performance.

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Section: Introductionmentioning

confidence: 99%

Nature of foulants and fouling mechanism in the Protein A MabSelect resin cycled in a monoclonal antibody purification process

Zhang

Daniels

Salm

et al. 2015

Biotech & Bioengineering

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“…An incubation time of 15 min was used and this corresponded to the CIP contact time in a column. Protocol for operating the HTPD chromatography platform was as has been suggested in the literature . Both single step and multi‐step CIP regimens were evaluated (Table ) .…”

Section: Resultsmentioning

confidence: 99%

“…With increase in urea concentration, ligand degradation increased from 10% to 18% while foulant clearance remained at ∼50%. Reducing agents aid removal of impurities by breaking the disulfide bonds . Use of DTT resulted in a foulant clearance of ∼60% with negligible ligand degradation.…”

Section: Resultsmentioning

confidence: 99%

“…Protocol for operating the HTPD chromatography platform was as has been suggested in the literature. 33,34 Both single step and multi-step CIP regimens were evaluated (Table 1). 33,35 -38 All experiments were performed in triplicate.…”

Section: Screening Of Cleaning In Place (Cip) Buffersmentioning

confidence: 99%

“…Reducing agents aid removal of impurities by breaking the disulfide bonds. 33 Use of DTT resulted in a foulant clearance of ∼60% with negligible ligand degradation. Among the three cleaning reagents examined in this study (NaOH, chaotropes and DTT), DTT was found to have a comparable foulant clearance capacity with that of NaOH, however, with negligible ligand degradation.…”

Section: Screening Of Cleaning In Place (Cip) Buffersmentioning

confidence: 99%

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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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Rapid Buffer and Ligand Screening for Affinity Chromatography by Multiplexed Surface Plasmon Resonance Imaging

Geuijen

Wijk-Basten

Egging

et al. 2017

Biotechnology Journal

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Protein purifications are often based on the principle of affinity chromatography, where the protein of interest selectively binds to an immobilized ligand. The development of affinity purification requires selecting proper wash and elution conditions. In recent years, miniaturization of the purification process is applied to speed up the development (e.g., microtiterplates, robocolumns). The application of surface plasmon resonance imaging (SPRi) as a tool to simultaneously screen many buffer conditions for wash and elution steps in an affinity-based purification process is studied. Additionally, the protein A ligand stability after exposure to harsh cleaning conditions often limits the reuse of resins and is determined at lab scale. The SPRi technology to screen ligand life-time with respect to alkali stability is used. It is also demonstrated that SPRi can successfully be applied in screening experiments for process developments in a miniaturized approach. The amount of resin, protein and buffer in these studies is reduced 30-300-fold compared to 1 mL column scale, and approximately 10-1000-fold compared to filter plate experiments. The overall development time can be decreased from several months towards days. The multiplexed SPRi can be applied in screening affinity chromatography conditions in early stage development for ligand development and recombinant protein production.

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A tool for increasing the lifetime of chromatography resins

Cited by 36 publications

References 2 publications

Nature of foulants and fouling mechanism in the Protein A MabSelect resin cycled in a monoclonal antibody purification process

Nature of foulants and fouling mechanism in the Protein A MabSelect resin cycled in a monoclonal antibody purification process

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

Rapid Buffer and Ligand Screening for Affinity Chromatography by Multiplexed Surface Plasmon Resonance Imaging

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