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Targeted CHO cell engineering approaches can reduce HCP‐related enzymatic degradation and improve mAb product quality
Author(s) -
Dovgan Tatiana,
Golghalyani Vahid,
Zurlo Fabio,
Hatton Diane,
Lindo Viv,
Turner Richard,
Harris Claire,
Cui Tingting
Publication year - 2021
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27857
Subject(s) - chinese hamster ovary cell , recombinant dna , downstream processing , cell culture , crispr , cas9 , microbiology and biotechnology , degradation (telecommunications) , upstream and downstream (dna) , rna interference , chemistry , biology , gene , biochemistry , rna , upstream (networking) , genetics , computer science , computer network , telecommunications
Host cell proteins (HCP) that co‐purify with biologics produced in Chinese hamster ovary cells have been shown to impact product quality through proteolytic degradation of recombinant proteins, leading to potential product losses. Several problematic HCPs can remain in the final product even after extensive purification. Each recombinant cell line has a unique HCP profile that can be determined by numerous upstream and downstream factors, including clonal variation and the protein sequence of the expressed therapeutic molecule. Here, we worked with recombinant cell lines with high levels of copurifying HCPs, and showed that in those cell lines even modest downregulation (≤50%) of the difficult to remove HCP Cathepsin D, through stable short hairpin RNA interference or monoallelic deletion of the target gene using CRISPR‐Cas9, is sufficient to greatly reduce levels of co‐purifying HCP as measured by high throughput targeted LC‐MS. This reduction led to improved product quality by reducing fragmentation of the drug product in forced degradation studies to negligible levels. We also show the potential of cell engineering to target other undesired HCPs and relieve the burden on downstream purification.