Fractionation and immunoreactivity of membrane associated proteins of Mycobacterium bovis AN5

Abstract

The membrane associated antigen was extracted from the pellet of M.hovis ANS sonicate with 2% SDS and 2%
mercaptoethanol. The extracted antigen was fractionated on gel filtration and anion exchange columns. The fractionated
regions were checked for their reactivity to anti M bovis ANS serum in ELISA. Among several fractionated regions obtained, the highly immunoreactive antigens were heterogenous polypeptides of 66 and 68 kDa and 26 and 28 kDa and a homogenolls polypeptide of 38 kDa. These 3 antigens were highly reactive in ELISA to anti M bovis· AN5 serum. The immunoblot analysis of crude membrane associated antigens revealed 6 reactive bands of 68,45,38.32,26,and 14 kDa. All the fractionated antigens showed cross- reactivity with other Mycobacterium species in ELISA.