Expression and purification of rEhCP112 and antibodies generation. (A) Expression and purification of rEhCP112. An Ehcp112 gene fragment (encoding the pro-peptide and the mature enzyme) cloned in the pTrcHIs C expression vector was expressed in E. coli. Then, the rEhCP112 was purified through Ni2+-affinity columns and analyzed by SDS-PAGE. Lane M, molecular weight markers; lane 1, uninduced bacteria extracts; lane 2, bacteria extracts induced by IPTG; lane 3, purified proteins corresponding to the pro-peptide and mature enzyme (52 and 43 kDa). (B) Antibodies generation. The rEhCP112 was inoculated in rabbits. Then, serum was tested by Western blot assays using the rEhCP112 in a SDS-PAGE. (C) Western blot on E. histolytica extracts. To analyze the specificity of the antibody against rEhCP112 we performed Western blot assays on total extracts of E. histolytica trophozoites; Lane 1, control using the preimmune serum; lane 2, antibody against rEhCP112.