Akta fplc chromatograph

Approximately 250 μg of GYL was subjected to size exclusion chromatography with a Superdex 75 Increase 10/30 GL (GE Healthcare, Uppsala, Sweden) equilibrated in 0.01 M TBS–Ca, pH 8.5, using an AKTA FPLC chromatograph (Amersham Pharmacia Biotech, Uppsala, Sweden). Protein was eluted with the same buffer at a flow rate of 0.5 mL/min, and the absorbance was monitored at 280 nm.

The purification of DENV2 virus particles was performed by ion exchange chromatography using the AKTA FPLC chromatograph (Amersham Pharmacia Biotech, Little Chalfont, UK) associated with a 1 mL HiTrapTM ANX (high sub) (GElifesciences, Boston, MA, USA) anion exchange column previously loaded with 60 mM sodium phosphate buffer (pH 7.2). A total of 10 mL of infected cell culture supernatant (VERO cell previously infected for 7 days with DENV2, MOI of 0.1) was used for purification. The elution procedure was carried out by applying a linear gradient from 60 to 600 mM of sodium phosphate (pH 7.2) in 15 column volumes (CV). The virus-containing fractions were combined, added with 5% sterile glycerol and stored at −80 ° C for later use.