The Use of a GroEL-BLI Biosensor to Rapidly Assess Preaggregate Populations for Antibody Solutions Exhibiting Different Stability Profiles

Pace SE, et al., 107(2):559-570, J Pharm Sci., 2018

Environmental stresses such as temperature, agitation, and light can cause protein-based drugs to aggregate to varying extents, thereby altering efficacy and safety. Described herein is the utility and pharmaceutical applicability of a previously reported chaperone GroEL-Bio Layer Interferometry (GroEL-BLI) based method to detect the formation of transiently formed, preaggregate species in therapeutic monoclonal antibody (mAb) samples. Four different therapeutic mAb candidates (2 IgG1 molecules, 1 IgG4 molecule, and 1 bispecific mAb) exposed to different environmental stresses were evaluated using an automated GroEL-BLI system. A Pall ForteBio Octet RED96 platform equipped with Streptavidin (SA) Biosensor probes was used to perform GroEL-BLI biosensor assays. Biotinylated GroEL was immobilized onto SA sensor tips. GroEL-SA biosensor tips were treated with BSA to minimize nonspecific binding of mAb to any unoccupied SA biosensor surfaces. Subsequently, the association and dissociation kinetics of antibodies (stressed or unstressed) were analyzed by dipping the GroEL loaded biosensor tips in various concentrations of antibody samples. Binding amplitudes (nm) at the association phase of each of the mAb samples were compared. ATP in an osmolyte mixture of urea and glycerol was used to remove bound antibody from GroEL. Overall results of this study suggest that this automated GroEL-BLI method can be used to rapidly identify preaggregate species in antibody solutions that exhibit different stability profiles.

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