Isolation of the cell wall-associated 34 kD antigen of paratuberculosis by electro elution

Abstract

Mycobacterium avium subsp. paratuberculosis is the causative agent of Jhone’s disease or paratuberculosis in ruminants. Effective control of paratuberculosis has been hampered by the lack of a specific antigen for diagnosis of paratuberculosis in early stages of infection. The 34 kD cell wall protein of MAP have been suggested as major species-specific immunodominant antigen in Jhone’s disease, but because of special interactions between MAP cell wall proteins the purification of 34 kD protein for further studies is difficult. In present study, a method has been developed for extraction of 34 kD antigen from SDS-PAGE gels. Following separation of extracted cell wall proteins by polyacrylamide gels, proteins were stained with coomassi blue and 34 kD protein isolated using electroelution. Then, salts and SDS were removed by dialysis and 34 kD antigen was concentrated by poly ethylene Glycol 6 kD. Analysis of the system efficiency showed high protein recovery from SDS-PAGE gels. The mentioned antigen may be candidate with potential for vaccine development against MAP infection. Key words: Paratuberculosis, 34 kD antigen, electro elution