Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography

Amit K. Dutta, Travis Tran, Boris Napadensky, Achyuta Teella, Gary Brookhart, Philip A. Ropp, Ada W. Zhang, Andrew D. Tustian, Andrew Zydney, Oleg Shinkazh

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an "after binder" to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (~0.67. g/L) and one with high titer (~6.9. g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products.

Original languageEnglish (US)
Pages (from-to)54-64
Number of pages11
JournalJournal of Biotechnology
Volume213
DOIs
StatePublished - Nov 10 2015

Fingerprint

Countercurrent Distribution
Monoclonal antibodies
Staphylococcal Protein A
Chromatography
Cell culture
Purification
Cell Culture Techniques
Monoclonal Antibodies
Proteins
Fluids
Antibodies
Productivity
Resins
Column chromatography
Cricetulus
Ovary
Binders
Pumps
Recovery
Kinetics

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Dutta, Amit K. ; Tran, Travis ; Napadensky, Boris ; Teella, Achyuta ; Brookhart, Gary ; Ropp, Philip A. ; Zhang, Ada W. ; Tustian, Andrew D. ; Zydney, Andrew ; Shinkazh, Oleg. / Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography. In: Journal of Biotechnology. 2015 ; Vol. 213. pp. 54-64.
@article{daea62c2637c45ccae435d8ff5ff0858,
title = "Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography",
abstract = "Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an {"}after binder{"} to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (~0.67. g/L) and one with high titer (~6.9. g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products.",
author = "Dutta, {Amit K.} and Travis Tran and Boris Napadensky and Achyuta Teella and Gary Brookhart and Ropp, {Philip A.} and Zhang, {Ada W.} and Tustian, {Andrew D.} and Andrew Zydney and Oleg Shinkazh",
year = "2015",
month = "11",
day = "10",
doi = "10.1016/j.jbiotec.2015.02.026",
language = "English (US)",
volume = "213",
pages = "54--64",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier",

}

Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography. / Dutta, Amit K.; Tran, Travis; Napadensky, Boris; Teella, Achyuta; Brookhart, Gary; Ropp, Philip A.; Zhang, Ada W.; Tustian, Andrew D.; Zydney, Andrew; Shinkazh, Oleg.

In: Journal of Biotechnology, Vol. 213, 10.11.2015, p. 54-64.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography

AU - Dutta, Amit K.

AU - Tran, Travis

AU - Napadensky, Boris

AU - Teella, Achyuta

AU - Brookhart, Gary

AU - Ropp, Philip A.

AU - Zhang, Ada W.

AU - Tustian, Andrew D.

AU - Zydney, Andrew

AU - Shinkazh, Oleg

PY - 2015/11/10

Y1 - 2015/11/10

N2 - Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an "after binder" to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (~0.67. g/L) and one with high titer (~6.9. g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products.

AB - Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an "after binder" to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (~0.67. g/L) and one with high titer (~6.9. g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products.

UR - http://www.scopus.com/inward/record.url?scp=84930218144&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84930218144&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2015.02.026

DO - 10.1016/j.jbiotec.2015.02.026

M3 - Article

VL - 213

SP - 54

EP - 64

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

ER -