2.6.6. Analytical size exclusion chromatography (A-SEC)
In order to further evaluate purity and size distribution of SXC
eluates, samples (250 μL at 158 μg/mL) were directly injected into a
Superose 6 Increase 10/300 GL column (29-0915-96, Cytiva, Sweden). All
runs were performed in PBS at a flow rate of 0.7 mL/min, and the
absorbance was monitored either at 280 or 620 nm using an ultra fast
liquid chromatography equipment (Shimadzu, Japan). The column was
calibrated using thyroglobulin (670 kD), γ-globulin (158 kD), ovalbumin
(44 kD), myoglobin (17 kD), and vitamin B12 (1.35 kD) (151-1901,
Bio-Rad, USA). Blue dextran with molecular mass of 2,000 kD (D5751,
Sigma-Aldrich, USA) was used to determine the void volume of the column.