Ovalbumin pH formulation study and multiple domain melting Tm deconvolution using MMS

Abstract

Identifying thermal stability of a protein is an essential part of therapeutic biologic development and formulation to ensure long shelf lives. However, existing techniques such as DSC and DSF lack structural insight associated with melting temperature (Tm), particularly for multi-domain proteins with multiple unfolding events.

In this study, Microfluidic Modulation Spectroscopy (MMS) is presented as a versatile alternative that simultaneously measures protein structure and stability. Using ovalbumin as a model protein, MMS identified optimal pH formulation conditions while resolving multiple structure-specific melting temperatures. In addition, MMS detected subtle higher-order structural changes following freeze-thaw stress, demonstrating its high sensitivity.

These results highlight MMS as a powerful tool for formulation development, enabling detailed, domain-level insight into protein stability that is not accessible with conventional thermal methods.