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Pragmatic mAb lead molecule engineering from a developability perspective
Author(s) -
Chi Bertie,
De Oliveira Geoffrey,
Gallagher Tom,
Mitchell Laura,
Knightley Louise,
Gonzalez Cynthia C.,
Russell Shan,
Germaschewski Volker,
Pearce Claire,
Sellick Christopher A.
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.27802
Subject(s) - context (archaeology) , computational biology , in silico , lead (geology) , mutagenesis , chemistry , biopharmaceutical , computer science , process (computing) , biochemical engineering , combinatorial chemistry , mutation , biology , biochemistry , genetics , paleontology , engineering , gene , operating system
Abstract As the number of antibody drugs being approved and marketed increases, our knowledge of what makes potential drug candidates a successful product has increased tremendously. One of the critical parameters that have become clear in the field is the importance of mAb “developability.” Efforts are being increasingly focused on simultaneously selecting molecules that exhibit both desirable biological potencies and manufacturability attributes. In the current study mutations to improve the developability profile of a problematic antibody that inconsistently precipitates in a batch scale‐dependent fashion using a standard platform purification process are described. Initial bioinformatic analysis showed the molecule has no obvious sequence or structural liabilities that might lead it to precipitate. Subsequent analysis of the molecule revealed the presence of two unusual positively charged mutations on the light chain at the interface of VH and VL domains, which were hypothesized to be the primary contributor to molecule precipitation during process development. To investigate this hypothesis, straightforward reversion to the germline of these residues was carried out. The resulting mutants have improved expression titers and recovered stability within a forced precipitation assay, without any change to biological activity. Given the time pressures of drug development in industry, process optimization of the lead molecule was carried out in parallel to the “retrospective” mutagenesis approach. Bespoke process optimization for large‐scale manufacturing was successful. However, we propose that such context‐dependent sequence liabilities should be included in the arsenal of in silico developability screening early in development; particularly since this specific issue can be efficiently mitigated without the requirement for extensive screening of lead molecule variants.