| Autor: |
Yuan G; Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States., Salipante PF; Materials Science and Engineering Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States., Hudson SD; Materials Science and Engineering Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States., Gillilan RE; Center for High-Energy X-ray Sciences at CHESS, Cornell University, Ithaca, New York 14853, United States., Huang Q; Center for High-Energy X-ray Sciences at CHESS, Cornell University, Ithaca, New York 14853, United States., Hatch HW; Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States., Shen VK; Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States., Grishaev AV; Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States., Pabit S; Analytical Enabling Capabilities, Merck & Co., Inc., Rahway, New Jersey 07065, United States., Upadhya R; Analytical Enabling Capabilities, Merck & Co., Inc., Rahway, New Jersey 07065, United States., Adhikari S; Analytical Enabling Capabilities, Merck & Co., Inc., Rahway, New Jersey 07065, United States., Panchal J; Sterile and Specialty Products, Merck & Co., Inc., Kenilworth, New Jersey 07033, United States., Blanco MA; Discovery Pharmaceutical Sciences, Merck & Co., Inc., West Point, Pennsylvania 19486, United States., Liu Y; Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States.; Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware 19716, United States. |
| Abstrakt: |
The solution viscosity and protein-protein interactions (PPIs) as a function of temperature (4-40 °C) were measured at a series of protein concentrations for a monoclonal antibody (mAb) with different formulation conditions, which include NaCl and sucrose. The flow activation energy ( E η ) was extracted from the temperature dependence of solution viscosity using the Arrhenius equation. PPIs were quantified via the protein diffusion interaction parameter ( k D ) measured by dynamic light scattering, together with the osmotic second virial coefficient and the structure factor obtained through small-angle X-ray scattering. Both viscosity and PPIs were found to vary with the formulation conditions. Adding NaCl introduces an attractive interaction but leads to a significant reduction in the viscosity. However, adding sucrose enhances an overall repulsive effect and leads to a slight decrease in viscosity. Thus, the averaged (attractive or repulsive) PPI information is not a good indicator of viscosity at high protein concentrations for the mAb studied here. Instead, a correlation based on the temperature dependence of viscosity (i.e., E η ) and the temperature sensitivity in PPIs was observed for this specific mAb. When k D is more sensitive to the temperature variation, it corresponds to a larger value of E η and thus a higher viscosity in concentrated protein solutions. When k D is less sensitive to temperature change, it corresponds to a smaller value of E η and thus a lower viscosity at high protein concentrations. Rather than the absolute value of PPIs at a given temperature, our results show that the temperature sensitivity of PPIs may be a more useful metric for predicting issues with high viscosity of concentrated solutions. In addition, we also demonstrate that caution is required in choosing a proper protein concentration range to extract k D . In some excipient conditions studied here, the appropriate protein concentration range needs to be less than 4 mg/mL, remarkably lower than the typical concentration range used in the literature. |
