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Residual on column host cell protein analysis during lifetime studies of protein A chromatography

Lintern, K; Pathak, M; Smales, CM; Howland, K; Rathore, A; Bracewell, DG; (2016) Residual on column host cell protein analysis during lifetime studies of protein A chromatography. Journal of Chromatography A , 1461 pp. 70-77. 10.1016/j.chroma.2016.07.055. Green open access

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Abstract

Capacity reduction in protein A affinity chromatography with extended cycling during therapeutic antibody manufacture is well documented. Identification of which residual proteins remain from previous cycles during the lifetime of these adsorbent materials is required to understand their role in this ageing process, but represents a significant metrological challenge. Scanning electron microscopy (SEM) and liquid chromatography mass spectrometry (LC–MS/MS) are combined to detect and map this phenomenon of protein carry-over. We show that there is a morphological change at the surface of the agarose resin, revealing deposits on the polymer fibres increasing with cycle number. The amount of residual host cell proteins (HCPs) by LC–MS/MS present on the resin is shown to increase 10-fold between 50 and 100 cycles. During this same period the functional class of the predominant HCPs associated with the resin increased in diversity, with number of proteins identified increasing 5-fold. This ageing is observed in the context of the product quality of the eluate HCP and protein A leachate concentration remaining constant with cycle number.

Type: Article
Title: Residual on column host cell protein analysis during lifetime studies of protein A chromatography
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.chroma.2016.07.055
Publisher version: http://dx.doi.org/10.1016/j.chroma.2016.07.055
Language: English
Additional information: © 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license http://creativecommons.org/licenses/by/4.0/).
Keywords: Science & Technology, Life Sciences & Biomedicine, Physical Sciences, Biochemical Research Methods, Chemistry, Analytical, Biochemistry & Molecular Biology, Chemistry, Protein A affinity chromatography, Host cell proteins, LC-MS/MS, Resin ageing, Process-related impurities, Bioprocessing, ANTIBODY PURIFICATION PROCESS, VIRUS-LIKE PARTICLES, MONOCLONAL-ANTIBODIES, IMMUNOGLOBULIN-G, IDENTIFICATION, PERFORMANCE, IMPURITIES, CLEARANCE, ECONOMICS, CHROMATIN
UCL classification: UCL
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
URI: https://discovery.ucl.ac.uk/id/eprint/1508915
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