Probing Microenvironmental Acidity in Lyophilized Protein and Vaccine Formulations Using Solid-state NMR Spectroscopy

Biophysical and biochemical instability of therapeutic proteins in the solution state may necessitate development products solid form, due to their enhanced stability. Lyophilization is a widely used method ensure dry stabilization biological products. A commonly encountered issue pH shifts that can occur undesired crystallization buffer component, resulting loss protein activities. However, it technically challenging noninvasively investigate physicochemical environment lyophile matrix. In this work, we demonstrate an approach based on solid-state NMR microenvironmental acidity lyophilized formulations, using histidine, agent, as molecular probe. The matrix be assessed by monitoring chemical shift changes histidine. protonation tautomeric states histidine at range values from 4.5 11.0 were identified full 13C 15N resonance assignments one-dimensional two-dimensional experiments. results demonstrated pH-dependence amorphous state. Moreover, successfully applied protocol formulations HPV vaccine lactate dehydrogenase protein.