PMID: 11344457May 10, 2001Paper

Isolation of a recombinant antibody from cell culture supernatant: continuous annular versus batch and expanded-bed chromatography

Biotechnology and Bioengineering
R Giovannini, Ruth Freitag

Abstract

Annular chromatography represents a crossflow approach to chromatographic separations, that allows the continuous separation of multicomponent mixtures. The potential of the method for continuous bioseparation has been discussed for some time, however, we demonstrate for the first time the processing of a complex feed (cell culture supernatant) taken from an actual (bio)process. Moreover, while previously published applications of annular chromatography concentrated on noninteractive (gel filtration) or nonspecific (ion exchange) chromatography, we show the possibility of continuous annular affinity chromatography. In particular, a commercially available preparative continuous annular chromatography (P-CAC) system was used to purify a recombinant antibody (human IgG(1)-kappa) from CHO cell culture supernatants by (pseudo)affinity chromatography on hydroxyapatite (HA) and rProtein A. Methods developed using small (2 mL) batch columns could be directly transferred to the P-CAC, where they yielded similar results in terms of final product quality. Yields were between 87% and 92% in the case of HA and between 77% and 82% in the case of rProtein A chromatography. DNA removal was nearly quantitative in all cases. Concomitantly, the a...Continue Reading

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Citations

Jul 9, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Stéphane CanarelliRuth Freitag
May 28, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Frank Hilbrig, Ruth Freitag
Jul 16, 2013·Trends in Biotechnology·Alois Jungbauer
Jul 28, 2015·Biotechnology Journal·Ruben R G SoaresM Raquel Aires-Barros
Jun 28, 2016·Materials·Vuk Uskoković, Victoria M Wu
Feb 13, 2015·Preparative Biochemistry & Biotechnology·Anurag S RathoreS Muthukumar
Dec 26, 2016·Methods : a Companion to Methods in Enzymology·Sushrut AroraB Vijayalakshmi Ayyar
Jan 12, 2002·Biotechnology and Bioengineering·R Giovannini, R Freitag
Jul 29, 2004·Biotechnology and Bioengineering·Alois JungbauerPing Luo
Oct 2, 2002·Biotechnology and Bioengineering·Jens H VogelKonstantin Konstantinov
Oct 31, 2009·Journal of Separation Science·Mark A SnyderPete Gagnon
Oct 31, 2009·Journal of Separation Science·Pete GagnonPaul J Yazaki
Aug 9, 2002·Journal of Agricultural and Food Chemistry·Berndt FinkeGuenther Boehm
Jun 6, 2009·New Biotechnology·Pete Gagnon

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