Identification and purification of a specific and immunogenic antigen of the laminated layer of the hydatid cyst and production of an antigen-specific monoclonal antibody

ABSTRACT

Cystic echinococcosis [CE] is an infection caused by the larval stage of Echinococcus granulosus. This is widely distributed through Iran, where a variety of animals act as intermediate host. The immunogenic antigens [Ag] of different compartments of the hydatid cyst have been already determined. One of these compartments is the laminated layer [LL]. We have extracted a protein with the MW of 24 kDa from a lysate prepared from the LL and produced a monoclonal antibody [mAb] against this protein. Five mAb named P[3]F[6s], P[2]Hp[4s], P[1], A[6s], P[1],C[3s], and P[1],F[7s] have been produced. The isotype analysis showed that P[3]F[6s] is IgG[1], and the rest are IgM. P[3]F[6s] was purified from the ascitic fluid of mice injected with P[3]F[6s] hybridoma intraperitoneally. Western blot and LLISA analysis showed that this mAb could recognize the purified 24 kDa prepared from lysate of LL. Since a 24 kDa protein has been shown to be an immunogenic Ag, this protein can be used as a candidate for the development of diagnostic tests and vaccine strategies. For these aims, P[3]F[6s] can be used for the purification of this 24 kDa protein