4.6. Gel Filtration Chromatography

Gel filtration chromatography was performed with the ÄKTA purifier protein purification system (GE Healthcare Life Sciences, Pittsburgh, PA, USA). A GE Filtration Calibration Kit HMW was used to calibrate the Superdex 200 10/300 increase column (GE Healthcare Life Sciences, Pittsburgh, PA, USA), and a standard curve was drawn based on five standard proteins: Thyroglobulin (669 kDa), Ferritin (440 kDa), Aldolase (158 kDa), Conalbumin (75 kDa), and Ovalbumin (44 kDa). All samples were centrifuged at 4 °C and 12,000× g for 20 min to remove the precipitated materials and soluble gases that were stored in the equilibration buffer (20 mM Tris, 300 mM NaCl, and 5% Glycerol at pH 7.5).