Continuous countercurrent tangential chromatography for mixed mode post-capture operations in monoclonal antibody purification

Amit K. Dutta, Dmitriy Fedorenko, Jasmine Tan, Joseph A. Costanzo, David S. Kahn, Andrew L. Zydney, Oleg Shinkazh

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Continuous Countercurrent Tangential Chromatography (CCTC) has been shown to demonstrate significant advantages over column chromatography including higher productivity, lower operational pressure, disposable flow path, and lower resin use. Previous applications of CCTC have been limited to initial capture of monoclonal antibodies (mAb) from clarified cell culture harvest. In this present article, a CCTC system was designed and tested for a post-capture antibody purification step. Mixed mode cation exchange-hydrophobic interaction chromatography resins with two different particle sizes were used to reduce host cell protein (HCP), leached protein A, DNA, and aggregates from a mAb stream after a protein A operation. Product output from CCTC was obtained at a steady-state concentration in sharp contrast to the periodic output of product in multi-column systems. The results show up to 101 g of mAb/L of resin/hr productivity, which is 10× higher than in a batch column. A 5% yield increase (95% with CCTC vs. 90% in batch column) resulted from optimizing elution pH within a narrow operational window (pH 4–4.5). Contaminant removal was found to be similar to conventional column performance. Data obtained with the smaller particle size resin showed faster binding kinetics leading to reduced CCTC system volume and increased productivity. Buffer and water usage were modeled to show potential for utilization of in-line mixing and buffer tank volume reduction. The experimental results were used to perform a scale up exercise that predicts a compact CCTC flow path for 500 and 2000 L batches using commercially available membranes. These results demonstrate the potential of using CCTC for post-capture operations as an alternative to packed bed chromatography, and provide a framework for the design and development of an integrated continuous bioprocessing platform based on CCTC technology.

Original languageEnglish (US)
Pages (from-to)37-44
Number of pages8
JournalJournal of Chromatography A
Volume1511
DOIs
StatePublished - Aug 18 2017

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Countercurrent Distribution
Chromatography
Purification
Monoclonal Antibodies
Resins
Staphylococcal Protein A
Particle Size
Productivity
Buffers
Particle size
Column chromatography
Hydrophobic and Hydrophilic Interactions
Packed beds
Cations
Cell culture
Cell Culture Techniques
Ion exchange
Technology
Pressure

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Dutta, Amit K. ; Fedorenko, Dmitriy ; Tan, Jasmine ; Costanzo, Joseph A. ; Kahn, David S. ; Zydney, Andrew L. ; Shinkazh, Oleg. / Continuous countercurrent tangential chromatography for mixed mode post-capture operations in monoclonal antibody purification. In: Journal of Chromatography A. 2017 ; Vol. 1511. pp. 37-44.
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abstract = "Continuous Countercurrent Tangential Chromatography (CCTC) has been shown to demonstrate significant advantages over column chromatography including higher productivity, lower operational pressure, disposable flow path, and lower resin use. Previous applications of CCTC have been limited to initial capture of monoclonal antibodies (mAb) from clarified cell culture harvest. In this present article, a CCTC system was designed and tested for a post-capture antibody purification step. Mixed mode cation exchange-hydrophobic interaction chromatography resins with two different particle sizes were used to reduce host cell protein (HCP), leached protein A, DNA, and aggregates from a mAb stream after a protein A operation. Product output from CCTC was obtained at a steady-state concentration in sharp contrast to the periodic output of product in multi-column systems. The results show up to 101 g of mAb/L of resin/hr productivity, which is 10× higher than in a batch column. A 5{\%} yield increase (95{\%} with CCTC vs. 90{\%} in batch column) resulted from optimizing elution pH within a narrow operational window (pH 4–4.5). Contaminant removal was found to be similar to conventional column performance. Data obtained with the smaller particle size resin showed faster binding kinetics leading to reduced CCTC system volume and increased productivity. Buffer and water usage were modeled to show potential for utilization of in-line mixing and buffer tank volume reduction. The experimental results were used to perform a scale up exercise that predicts a compact CCTC flow path for 500 and 2000 L batches using commercially available membranes. These results demonstrate the potential of using CCTC for post-capture operations as an alternative to packed bed chromatography, and provide a framework for the design and development of an integrated continuous bioprocessing platform based on CCTC technology.",
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Continuous countercurrent tangential chromatography for mixed mode post-capture operations in monoclonal antibody purification. / Dutta, Amit K.; Fedorenko, Dmitriy; Tan, Jasmine; Costanzo, Joseph A.; Kahn, David S.; Zydney, Andrew L.; Shinkazh, Oleg.

In: Journal of Chromatography A, Vol. 1511, 18.08.2017, p. 37-44.

Research output: Contribution to journalArticle

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