A streamlined workflow for early phase formulation development of monoclonal antibodies comprising multi-attribute method and ligand binding assay.

A streamlined early phase formulation development workflow has been developed for monoclonal antibodies (mAbs) to provide a more efficient process, driving down costs and reducing timelines, without compromising the quality and hence patient safety. The proposed novel workflow combines liquid chromatography-mass spectrometry multi-attribute method (LC-MS MAM) and sensitive ligand binding using surface plasmon resonance (SPR). By linking the two methodologies it is possible to obtain a comprehensive understanding of a mAb's critical quality attributes (CQAs) and provide a structure/function correlation. As LC-MS MAM cannot address all aspects of degradation, high throughput methods for the analysis of high molecular weight material (HMWM), and conformational and colloidal stability, were also evaluated. The workflow comprises an initial forced degradation study, to verify stability-indication and identify potential degradation routes. Secondly, optimal pH, based on conformational and colloidal stability, is determined. Finally, stabilising excipients are evaluated by design of experiment (DoE). We have verified this workflow using pembrolizumab. In an initial forced degradation study, LC-MS MAM and PD-1 ligand binding could identify the CQAs. Met105 oxidation, located in the CDR3 region, was identified as the major CQA. DoE demonstrated that 25 mM methionine inhibited Met105 oxidation and stabilised PD-1 binding. With this streamlined process, we were able to improve the stability of the protein by formulating in 20 mM histidine, 25 mM methionine, 0.02 % PS80 and 300 mM sucrose, at pH 5.5. The described workflow has the potential to decrease the demand for precious early development material as well as reduce costs and shorten timelines.