Investigating the Correlative Nature of Calcitonin Aggregation and Solvent Evaporation Kinetics

• Jordan Gold, Biochemistry, University of Delaware

• Ariel Alperstein, Chemistry and Biochemistry, University of Delaware

Protein misfolding and aggregation are characteristic features of amyloid-associated diseases. Current methods to detect protein aggregation require a combination of imaging, spectroscopic, and fluorescence assays. Here, we develop a new benchtop method that investigates the correlation between solvent evaporation and protein aggregation.  Human calcitonin (hCT) is a 32-amino acid hormonal amyloidogenic peptide that serves as an ideal model for this investigation.  Samples were incubated in multiple environments, and evaporation was measured using an analytical balance. Protein aggregation and kinetics were confirmed and monitored using Thioflavin T (ThT) fluorescence assays. Furthermore, aggregate morphology and secondary structure were determined using Transmission Electron Microscopy (TEM) and Fourier Transform Infrared Spectroscopy (FTIR).  Early findings indicate that these processes are correlative, showing a direct relationship between aggregation kinetics and solvent evaporation. These findings suggest that evaporation is an important variable influencing the rate and presence of aggregation. Further understanding this relationship can provide the field with new methods and improve experimental design for protein aggregation studies.