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

Lintern, Katherine, Pathak, Mili, Smales, Christopher Mark, Howland, Kevin, Rathore, Anurag, Bracewell, Daniel G. (2016) Residual on column host cell protein analysis during lifetime studies of protein A chromatography. Journal of Chromatography A, 1461 . pp. 70-77. ISSN 0021-9673. (doi:10.1016/j.chroma.2016.07.055) (KAR id:56828)

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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.

Item Type: Article
DOI/Identification number: 10.1016/j.chroma.2016.07.055
Uncontrolled keywords: Protein A affinity chromatography; Host cell proteins; LC–MS/MS; Resin ageing; Process-related impurities; Bioprocessing
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > School of Biosciences
Depositing User: Mark Smales
Date Deposited: 12 Aug 2016 15:23 UTC
Last Modified: 29 May 2019 17:42 UTC
Resource URI: (The current URI for this page, for reference purposes)
Smales, Christopher Mark:
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