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Identification of Upstream Culture Conditions and Harvest Time Parameters That Affect Host Cell Protein Clearance

Wilson, LJ; Lewis, W; Kucia-Tran, R; Bracewell, DG; (2019) Identification of Upstream Culture Conditions and Harvest Time Parameters That Affect Host Cell Protein Clearance. Biotechnology Progress , 35 (3) , Article e2805. 10.1002/btpr.2805. Green open access

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Abstract

During early stage bioprocess development, characterizing interactions between unit operations is a key challenge. Such interactions include the release of host cell enzymes early in the process causing losses in product quality downstream. Using a CHO-expressed IgG1 system, the impact of cell culture duration was investigated using a 50 L bioreactor and performing scale-down protein A purification. While antibody titer doubled during the last week of culture, the post-protein A HCP levels increased from 243 to 740 ppm. Effects of pH and temperature were then explored using fed-batch ambr250 bioreactors, and parameters enabling higher titers were linked to a decrease in post-protein A product purity. These trade-offs between titer and product quality were visualised using a window of operation. The downstream space was explored further by exposing shake flask material to shear representative of disc stack centrifugation, prior to purification, and by adding polishing chromatography. While product quality decreased with progressing cultivation, cells became more shear resistant. Polishing chromatography resulted in product fragmentation which increased 4-fold from day 10 to 24, adding constraint to achieving both efficient HCP clearance as well as high monomer purities. These examples highlight the importance of adopting integrated approaches to upstream and downstream development strategies to enable whole process optimisation. This article is protected by copyright. All rights reserved.

Type: Article
Title: Identification of Upstream Culture Conditions and Harvest Time Parameters That Affect Host Cell Protein Clearance
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/btpr.2805
Publisher version: http://doi.org/10.1002/btpr.2805
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: downstream processing, host cell proteins, monoclonal antibodies, product quality, upstream processing
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/10070543
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