Pragmatic mAb lead molecule engineering from a developability perspective.

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 has 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 is 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 hypothesised to be the primary contributor to molecule precipitation during process development. To investigate this hypothesis, straightforward reversion to germline of these residues was carried out. The resulting mutants have improved expression titres and recovered stability within a forced precipitation assay, without any change to biological activity. Given the time pressures of drug development in industry, process optimisation of the lead molecule was carried out in parallel to the 'retrospective' mutagenesis approach. Bespoke process optimisation for large-scale manufacture 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. This article is protected by copyright. All rights reserved.