Figure 4From: Purification of native HBHA from Mycobacterium avium subsp. paratuberculosisHeparin-Sepharose chromatography of a Map lysate in PBS buffer. A Map K-10 culture obtained using the bioreactor was centrifuged and the pellet was resuspended in 0.5 × PBS. The bacteria were then sonicated and centrifuged. The whole cell lysate (WCL) was applied onto a heparin-Sepharose HiTrap column and eluted using a 0-1 M NaCl gradient. WCL and the indicated elution fractions were analyzed by SDS-PAGE. Fractions 20 to 25, eluted around 500 mM NaCl, contained map-Lbp/Hlp, as determined HR-MS/MS. The molecular weights (MW) expressed in kDa are indicated in the left margin.Back to article page