Figure 1

Purification of rSjB6 expressed in Hi-5 cells by IMAC affinity chromatography followed by size exclusion chromatography. Top panel: (A) An IMAC column was loaded with filtered protein sample and the column washed to remove unbound material. (B) Multi-step elution was carried out with increasing concentration of elution buffer (4%, 8%, 16%, 26%, 50% and 100%). The peaks indicate eluted protein fractions. (C) IMAC purified elution samples were pooled, concentrated and loaded onto the SEC column for further purification. The blue peak corresponds to the rSjB10 fraction. Bottom panel: Representative samples collected from the eluted fractions were analysed by SDS-PAGE. L = protein ladder, lane 1 = culture supernatant; lane 2 = filtered supernatant; lane 3 = Crossflow waste; lane 4 = concentrated supernatant; lane 5 = IMAC flow through; lane 6 = IMAC wash. Black arrows show the purified rSjB6.