Two-stage chromatographic separation of aggregates for monoclonal antibody therapeutics

Kumar, Vijesh; Rathore, Anurag S.

doi:10.1016/j.chroma.2014.09.077

Cited by 17 publications

(10 citation statements)

References 22 publications

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“…Cation exchange chromatography in bind‐elute mode is the most widely used method for profiling the charge heterogeneity of monoclonal antibodies . Hydrophobic interaction (HIC), anion exchange (AEX), and multimodal chromatography (MMC) in bind‐elute and flow through modes have been used by many researchers for removal of HMWI . In a recent study, a two stage chromatographic method comprising of CEX in bind‐elute and HIC in flow‐through mode was shown to achieve complete removal of HMWI from mAb process streams .…”

Section: Resultsmentioning

confidence: 99%

“…Hydrophobic interaction (HIC), anion exchange (AEX), and multimodal chromatography (MMC) in bind‐elute and flow through modes have been used by many researchers for removal of HMWI . In a recent study, a two stage chromatographic method comprising of CEX in bind‐elute and HIC in flow‐through mode was shown to achieve complete removal of HMWI from mAb process streams . With the motivation to develop an integrated process for simultaneous separation of charge variants and aggregates, the possibility of using a two‐step process (CEX followed by MMC) was explored such as the CEX in bind‐elute mode could be used to target separation of the charge variants and MMC in flow through mode for HMWI removal.…”

Section: Resultsmentioning

confidence: 99%

“…CEX and MMC steps were optimized individually followed by their integration. Resin media with a bead size of ≥50 µm and resin bed heights of ≤20 cm were used to avoid high back pressure . Detailed optimization studies were carried out for mAb C and the data are presented in the following sections.…”

Section: Resultsmentioning

confidence: 99%

“…Cumulative plots for HMWI are routinely used to analyze the performance of a resin with regard to resolution of HMWI . Closer is the curve to X‐axis, better is the performance.…”

Section: Methodsmentioning

confidence: 99%

“…This platform has been shown to perform consistently for multiple mAbs, delivering product containing high molecular weight impurities (HMWI) of <1% and HCP of <50 ppm. In another recent development, an integrated two‐stage platform process for separation of aggregates has been proposed and offers near complete separation of aggregates with >95% recovery and reduced process time . As seen from above discussion, most of the reported developments have been focussed on development of mAb platform processes for separation of aggregates.…”

Section: Introductionmentioning

confidence: 99%

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Integrated Chromatographic Platform for Simultaneous Separation of Charge Variants and Aggregates from Monoclonal Antibody Therapeutic Products

Kateja

Kumar

Godara

et al. 2017

Biotechnology Journal

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Achieving consistent product quality of a biotherapeutic is a major target for any biopharmaceutical manufacturer, even more for a biosimilar producer as comparability with the innovator product is a regulatory expectation. The complexity of biotherapeutic products and their tedious manufacturing processes, however, make this a non-trivial exercise. The primary motivation of this work is to develop an integrated chromatographic platform for purification of monoclonal antibody (mAb) therapeutics that can deliver the desired separation of both charge variants and aggregates, in addition to the process related impurities like host cell proteins (HCP) and host cell DNA. To achieve the same, an integrated two-stage chromatographic process platform consisting of cation exchange chromatography and multimodal chromatography is being proposed. The versatility of the proposed platform has been successfully demonstrated for three different mAbs. It have been shown that in each case charge variant separation is achieved with the required clearance of aggregates (<1%), HCP (<10 ppm), and DNA (<5 ppb). Moreover, the proposed platform is conducive to use for development of a continuous process and offers smaller process time, lower buffer utilization, and decreased operational costs when compared to the conventional purification platforms.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…Cumulative plots for HMWI are routinely used to analyze the performance of a resin with regard to resolution of HMWI . Closer is the curve to X‐axis, better is the performance.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Integrated Chromatographic Platform for Simultaneous Separation of Charge Variants and Aggregates from Monoclonal Antibody Therapeutic Products

Kateja

Kumar

Godara

et al. 2017

Biotechnology Journal

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Hydrophobic Interaction Chromatography for the Purification of Antibodies

Vajda¹,

Müller²

2017

Process Scale Purification of Antibodies

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Integrated continuous processing of proteins expressed as inclusion bodies: GCSF as a case study

Kateja

Agarwal

Hebbi

et al. 2017

Biotechnology Progress

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Affordability of biopharmaceuticals continues to be a challenge, particularly in developing economies. This has fuelled advancements in manufacturing that can offer higher productivity and better economics without sacrificing product quality in the form of an integrated continuous manufacturing platform. While platform processes for monoclonal antibodies have existed for more than a decade, development of an integrated continuous manufacturing process for bacterial proteins has received relatively scant attention. In this study, we propose an end-to-end integrated continuous downstream process (from inclusion bodies to unformulated drug substance) for a therapeutic protein expressed in Escherichia coli as inclusion body. The final process consisted of a continuous refolding in a coiled flow inverter reactor directly coupled to a three-column periodic counter-current chromatography for capture of the product followed by a three-column con-current chromatography for polishing. The continuous bioprocessing train was run uninterrupted for 26 h to demonstrate its capability and the resulting output was analyzed for the various critical quality attributes, namely product purity (>99%), high molecular weight impurities (<0.5%), host cell proteins (<100 ppm), and host cell DNA (<10 ppb). All attributes were found to be consistent over the period of operation. The developed assembly offers smaller facility footprint, higher productivity, fewer hold steps, and significantly higher equipment and resin utilization. The complexities of process integration in the context of continuous processing have been highlighted. We hope that the study presented here will promote development of highly efficient, universal, end-to-end, fully continuous platforms for manufacturing of biotherapeutics. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:998-1009, 2017.

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Two-stage chromatographic separation of aggregates for monoclonal antibody therapeutics

Cited by 17 publications

References 22 publications

Integrated Chromatographic Platform for Simultaneous Separation of Charge Variants and Aggregates from Monoclonal Antibody Therapeutic Products

Integrated Chromatographic Platform for Simultaneous Separation of Charge Variants and Aggregates from Monoclonal Antibody Therapeutic Products

Hydrophobic Interaction Chromatography for the Purification of Antibodies

Integrated continuous processing of proteins expressed as inclusion bodies: GCSF as a case study

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