Pressure-perturbation of protein secondary structure coupled with Microfluidic Modulation Spectroscopy – a powerful platform for biopharmaceutical formulations development
November 6, 2020
Biophysical Journal | February 7, 2020
Vera Gross, Gary Smejkal, Nicole Cutri, Alexander Lazarev (Pressure Biosciences) Libo Wang, John Linnan, Matthew McGann, Jeffrey Zonderman, (RedShiftBio)
Biophysical Journal, vol. 118, issue 3, p. 37a
High hydrostatic pressure is a thermodynamic driver causing unfolding of proteins orthogonal to the action of temperature or various chaotropic reagents. Pressure effects on protein conformation are explained by hydration of solvent-excluded cavities that are populated with solvent upon unfolding.
Infrared spectroscopy, together with circular dichroism and fluorescence, is a popular methods of monitoring protein structure changes. Recently introduced Microfluidic Modulation Spectroscopy (MMS) represents a major advancement of infrared spectroscopy specifically developed to simplify protein structure analysis. Pressure effects on proteins is highly reproducible and can be controlled very precisely. Pressure perturbation approach coupled with MMS can be used to study stability of human immunoglobulins as a model system for formulations development of monoclonal antibody products and other biopharmaceuticals.
In this study, we demonstrate that pressure unfolding of human immunoglobulins in specific chemical environments promotes quantitative conversion of parallel beta sheet structures to the anti-parallel beta structures, a characteristic indicator of amyloid protein aggregation. We explore pressure effects on aggregation kinetics in a series of co-solvents, chaotropes and popular stabilizing excipients. Pressure perturbation approach coupled with MMS can be used to study stability of human immunoglobulins as a model system for formulations development of monoclonal antibody products and other biopharmaceuticals.