2.3. High performance size exclusion chromatography
High performance size exclusion chromatography (HP-SEC) was applied for
evaluating the capability of 2D-AEC×SEC for production NSP-free FMD
vaccine, as well as the most important factors affecting the quality of
the final products, integrity of active ingredient of vaccine, by
monitoring fingerprint HPLC elution profiles of the purified FMDV
fractions throughout the purification procedure as describe before
(Bagheri et al., 2018). Briefly, HP-SEC analysis was performed on a
Knauer HPLC system equipped with a photodiode array detector (PDA) set
up at 280 nm and controlled by EZChrom Elite software. The HPLC
conditions included Ultrahydrogel Linear (300 × 7.8 mm, I.D.) analytical
column from Waters (NJ 07054, US), solvent system consisting of 50 mM
phosphate buffer with pH 7.2 containing 100 mM
Na2SO4 as the mobile phase, the flow
rate of 0.6 mL/min, and the injection volume of 100 µl.