Purification of recombinant gp120 protein

Supernatants containing MGAT1⁻ CHO expressed TZ97008 rgp120 were concentrated and buffer exchanged to 10 mM TRIS, pH = 8.0 buffer (Vivaflow 200 Sartorius, Göttingen, Germany). Eluent was monitored at the 280 nm wavelength using an Akta purifier (GE healthcare Chicago, IL). Concentrated and buffer-exchanged MGAT1⁻ CHO supernatant containing TZ97008 rgp120 was passed through a 5 cm anion exchange (HiTrap Q FF column, GE Healthcare, Chicago, IL) and flow through fractions containing gp120 were collected. These fractions were further purified via size exclusion chromatography using a 60 cm Highload Superdex 200pg column (GE Healthcare) in pH 8 TBS (Tris-buffered Saline) buffer. CHO-S expressed TZ97008 rgp120 was purified by affinity chromatography against the N-terminal gD tag, followed by size exclusion chromatography, as previously described (9 (link)).