pH‐based cation exchange chromatography in the capture and elution of monoclonal antibodies

Ng, Phillip; Snyder, Mark A.

doi:10.1002/jssc.201100720

Cited by 10 publications

(7 citation statements)

References 19 publications

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“…IEX is the preferred technique for preparative and process scale separation for charge variants. While bench‐top analytical scale resolution has improved tremendously (Amand et al, ; Farnan & Moreno, ; Fekete et al, , ; Follman & Fahrner, ; Joshi, Kumar, & Rathore, ; Kang, Kutzko, Hayes, & Frey, ; Lingg et al, ; Ng & Snyder, ; Nordborg et al, ; Rea, Moreno, Lou, & Farnan, ; Santora, Kaymakcalan, Sakorafas, Krull, & Grant, ; Talebi et al, ; Talebi, Shellie, Hilder, Lacher, & Haddad, ; Tao, Carta, Ferreira, & Robbins, , ; Tao, Perez, Carta, Ferreira, & Robbins, ; Weitzhandler et al, ; Zhang, Patapoff, Farnan, & Zhang, ), industrial separation of charge variants remains challenging due to the scale and capacity required. For most industrial processes, the stationary phase, physical parameters of the columns, are already fixed.…”

Section: Separation and Purification Of Charge Variants In Downstreammentioning

confidence: 99%

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Chung

Tian

Tan

et al. 2018

Biotech & Bioengineering

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Controlling the charge profile of therapeutic protein is a critical challenge in the current quality-by-design (QbD) paradigm, throughout all phases of biologics process development (PD): cell line development, upstream cell culture, recovery process, downstream purification, and analytical characterization. Charge variant profiles may influence efficacy and/or lead to unintended side-effects. Thus, maintaining a consistent charge profile is of tremendous importance, and increasingly, researchers have focused efforts toward developing strategies to mitigate variability during cell culture and to improve separation and detection of charge variants. Current understanding of factors affecting charge variant formation during manufacturing remains inadequate, and sometimes, even substantial commitment of resources may still not fully achieve the desired or consistent profiles. As such, this review attempts to provide a comprehensive resource for the biologics community by summarizing the impact of charge variants and CQA management, analytical methods for charge variant detection, as well as strategies in downstream and upstream PD for controlling charge variant profiles.

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Section: Separation and Purification Of Charge Variants In Downstreammentioning

confidence: 99%

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Chung

Tian

Tan

et al. 2018

Biotech & Bioengineering

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“…Abbreviations: ATPS, aqueous two-phase system; ATPE, aqueous two-phase extraction; CHO, Chinese hamster ovary; HPS, hydroxypropyl starch; IgG, immunoglobulin G; mAb, monoclonal antibody; VR, volume ratio regeneration, ligand leaching, and antibody aggregation at low pH elution conditions [6]. Recently, a variety of alternative techniques have been developed to improve the process efficiency and economics of antibody production, including mixed-mode adsorption [7], membrane adsorption [8], cationexchange chromatography [6], fluoroapatite chromatography [9], and aqueous two-phase extraction (ATPE).…”

Section: Introductionmentioning

confidence: 99%

“…Recently, a variety of alternative techniques have been developed to improve the process efficiency and economics of antibody production, including mixed-mode adsorption [7], membrane adsorption [8], cationexchange chromatography [6], fluoroapatite chromatography [9], and aqueous two-phase extraction (ATPE).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a PEG/hydroxypropyl starch aqueous two‐phase system for the separation of monoclonal antibodies from cell culture supernatant

Lin

Zhang

et al. 2014

J of Separation Science

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In this study, an aqueous two-phase system (ATPS) with PEG and hydroxypropyl starch (HPS) was used to separate monoclonal antibody (mAb) from Chinese hamster ovary cell culture supernatant. The phase diagram of the PEG/HPS ATPS was determined, and the effects of NaCl addition were investigated. The results showed that NaCl addition could lead to a shift of the binodal curve and that phase separation would occur at higher PEG and HPS concentrations. The effects of NaCl addition, pH, and the load of cell supernatant on the partitioning of mAb in a PEG/HPS ATPS were investigated. It was found that with 6% cell supernatant and 15% NaCl addition at pH 6.0, the yield of mAb in the upper phase was 96.7% with a purity of 96.0%. The back-extraction of mAb with a PEG/phosphate ATPS were also studied, and the results showed that after the two-step extraction with ATPSs the purity of mAb could reach 97.6 ± 0.5% with a yield of 86.8 ± 1.0%, which was comparable to the purification with Protein A chromatography. These results indicate that the two-step extraction with PEG/HPS and PEG/phosphate ATPSs might be a promising alternative for the separation of mAb from cell culture supernatant.

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“…These include low‐binding capacity, high cost, lack of an effective cleaning procedure, or short lifetime. Nonaffinity capture methods (e.g., ion exchange and mixed mode) have been evaluated in several studies and separation of HCPs is often found to be challenging because of the nonspecific nature of these separation techniques . Therefore, an in‐depth understanding of the HCP chromatographic behavior would provide valuable insight for the rational design of purification processes …”

Section: Introductionmentioning

confidence: 99%

“…Nonaffinity capture methods (e.g., ion exchange and mixed mode) have been evaluated in several studies and separation of HCPs is often found to be challenging because of the nonspecific nature of these separation techniques. [14][15][16][17] Therefore, an in-depth understanding of the HCP chromatographic behavior would provide valuable insight for the rational design of purification processes. 18 Besides the immunogenicity concern, HCPs can also compromise product quality and shelf-life by resulting in degradation of protein or excipient, which leads to fragmentation, aggregation, and particle formation.…”

Section: Introductionmentioning

confidence: 99%

Cathepsin L Causes Proteolytic Cleavage of Chinese‐Hamster‐Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

Luo¹,

Liu²,

Cao³

et al. 2018

Biotechnology Progress

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A stochastic approach of copurification of the protease Cathepsin L that results in product fragmentation during purification processing and storage is presented. Cathepsin L was identified using mass spectroscopy, characterization of proteolytic activity, and comparison with fragmentation patterns observed using recombinant Cathepsin L. Cathepsin L existed in Chinese hamster ovary cell culture fluids obtained from cell lines expressing different products and cleaved a variety of recombinant proteins including monoclonal antibodies, antibody fragments, bispecific antibodies, and fusion proteins. Therefore, characterization its chromatographic behavior is essential to ensure robust manufacturing and sufficient shelf life. The chromatographic behaviors of Cathepsin L using a variety of techniques including affinity, cation exchange, anion exchange, and mixed mode chromatography were systematically evaluated. Our data demonstrates that copurification of Cathepsin L on nonaffinity modalities is principally because of similar retention on the stationary phase and not through interactions with product. Lastly, Cathespin L exhibits a broad elution profile in cation exchange chromatography (CEX) likely because of its different forms. Affinity purification is free of fragmentation issue, making affinity capture the best mitigation of Cathepsin L. When affinity purification is not feasible, a high pH wash on CEX can effectively remove Cathepsin L but resulted in significant product loss, while anion exchange chromatography operated in flow‐through mode does not efficiently remove Cathepsin L. Mixed mode chromatography, using Capto™ adhere in this example, provides robust clearance over wide process parameter range (pH 7.7 ± 0.3 and 100 ± 50 mM NaCl), making it an ideal technique to clear Cathepsin L. © 2018 American Institute of Chemical Engineers Biotechnol. Prog ., 35: e2732, 2019

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pH‐based cation exchange chromatography in the capture and elution of monoclonal antibodies

Cited by 10 publications

References 19 publications

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Evaluation of a PEG/hydroxypropyl starch aqueous two‐phase system for the separation of monoclonal antibodies from cell culture supernatant

Cathepsin L Causes Proteolytic Cleavage of Chinese‐Hamster‐Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

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