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

Journal of Biotechnology
Amit K DuttaOleg Shinkazh

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 chromat...Continue Reading

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Citations

Jul 15, 2015·Biotechnology and Bioengineering·Andrew L Zydney
Jul 5, 2016·Biotechnology Journal·Fabian SteinebachMassimo Morbidelli
Sep 3, 2017·Biotechnology and Bioengineering·Anirudh M K NambiarAndrew L Zydney
Apr 28, 2018·Biotechnology Letters·Anurag S RathoreNikhil Kateja
May 31, 2017·Biotechnology Progress·Michael D HughsonLeda R Castilho
Dec 1, 2019·Biotechnology and Bioengineering·Dmitriy FedorenkoOleg Shinkazh
Jan 11, 2020·Frontiers in Bioengineering and Biotechnology·Nagesh K Tripathi, Ambuj Shrivastava
Mar 30, 2019·Journal of Pharmaceutical Innovation·Ou YangMarianthi Ierapetritou
Apr 13, 2021·MAbs·Ohnmar Khanal, Abraham M Lenhoff

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