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Identification of upstream culture conditions and harvest time parameters that affect host cell protein clearance
Author(s) -
Wilson Louisa J.,
Lewis Will,
KuciaTran Richard,
Bracewell Daniel G.
Publication year - 2019
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.2805
Subject(s) - bioreactor , bioprocess , downstream processing , titer , chromatography , chemistry , centrifugation , cross flow filtration , biology , antibody , biochemistry , immunology , paleontology , organic chemistry , membrane
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 host cell protein (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 visualized 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 fourfold 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 optimization.