Physical-chemical methodologies for molecular characterization of carrier proteins used in Bio-Manguinhos conjugated vaccines

Abstract

Methodology: SEC chromatographic profile of two different samples of TT (TT1 and TT2) were performed using three different columns (Zorbax®GF450, TSK®G4000 SWXL and SuperdexTM 200). Electrophoretic techniques such as SDS-PAGE, IEF-PAGE and NATIVE-PAGE were used to evaluate electrophoretic profile. Stability of secondary and tertiary structures were analyzed by circular dichroism and fluorescence spectroscopy respectively, with temperature varying between 25°C-85°C. All proteins were subjected to tryptic hydrolysis aiming obtain a peptide map by RPC chromatography. TT1 was used as standard for all analysis.