Fig. 3.

Purification of rPstS-1. a SDS-PAGE 12% during purification process. b Immunodetection of protein obtained by ultrafiltration and RP-HPLC. c Separation of components obtained after tangential filtration in a XBridge Protein BEH C4 reverse-phase column, from which the component eluting at 20 min was shown to be homogeneous. The gradient started by 5 min at 20% of B, then separation was run from 20 to 60% solution B, over next 20 min, and the pure rPstS-1 eluted at 20.74 min. Lanes M: molecular weight marker; 1, total protein from supernatant of the 27 h growth in BMGY; 2, total protein from supernatant obtained after 72 h under induction; 3, 10–100 kDa protein cut off from tangential filtration; 4, protein purify by RP-HPLC