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Fouling of an anion exchange chromatography operation in a monoclonal antibody process: Visualization and kinetic studies

Close, EJ; Salm, JR; Iskra, T; Sørensen, E; Bracewell, DG; (2013) Fouling of an anion exchange chromatography operation in a monoclonal antibody process: Visualization and kinetic studies. Biotechnology and Bioengineering , 110 (9) pp. 2425-2435. 10.1002/bit.24898. Green open access

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Abstract

Fouling of chromatographic resins over their operational lifetimes can be a significant problem for commercial bioseparations. In this article, scanning electron microscopy (SEM), batch uptake experiments, confocal laser scanning microscopy (CLSM) and small-scale column studies were applied to characterize a case study where fouling had been observed during process development. The fouling was found to occur on an anion exchange (AEX) polishing step following a protein A affinity capture step in a process for the purification of a monoclonal antibody. Fouled resin samples analyzed by SEM and batch uptake experiments indicated that after successive batch cycles, significant blockage of the pores at the resin surface occurred, thereby decreasing the protein uptake rate. Further studies were performed using CLSM to allow temporal and spatial measurements of protein adsorption within the resin, for clean, partially fouled and extensively fouled resin samples. These samples were packed within a miniaturized flowcell and challenged with fluorescently labeled albumin that enabled in situ measurements. The results indicated that the foulant has a significant impact on the kinetics of adsorption, severely decreasing the protein uptake rate, but only results in a minimal decrease in saturation capacity. The impact of the foulant on the kinetics of adsorption was further investigated by loading BSA onto fouled resin over an extended range of flow rates. By decreasing the flow rate during BSA loading, the capacity of the resin was recovered. These data support the hypothesis that the foulant is located on the particle surface, only penetrating the particle to a limited degree. The increased understanding into the nature of the fouling can help in the continued process development of this industrial example.

Type: Article
Title: Fouling of an anion exchange chromatography operation in a monoclonal antibody process: Visualization and kinetic studies
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/bit.24898
Publisher version: http://dx.doi.org/10.1002/bit.24898
Language: English
Additional information: © 2013 Wiley Periodicals, Inc. This work is licensed under the Creative Commons Attribution License, Attribution 3.0 Unported (CC BY 3.0) http://creativecommons.org/licenses/by/3.0/ which permits unrestricted use, distribution, and reproduction in any medium. However, you must attribute the work to the author (but not in any way that suggests that they endorse you or your use of the work).
Keywords: Chromatography resin fouling, Scanning electron microscopy, Confocal laser scanning microscopy, Anion exchange chromatography, Therapeutic protein
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Chemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/1389468
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