[Position analysis of three recombinant proteins of Echinococcus granulosus protoscolex with two-dimensional electrophoresis].

ABSTRACT

OBJECTIVE: To obtain specific antibodies of the three recombinant antigens obtained previously, rEgZW-5, rEg14-3-3 and rEgP-29, for identifying the corresponding proteins in two-dimensional electrophoretogram of Echinococcus granulosus protoscolex. METHODS: The distribution of proteins from E. granulosus protoscoleces was judged by SDS-PAGE previously. Two-dimensional electrophoresis was used to separate proteins from E. granulosus protoscoleces, and the result was scanned and analyzed by the PDquest software to get the information about the quantity of proteins as well as their isoelectric point (IP) and relative molecular mass (MA,). Rabbits were immunized with the 3 recombinant antigens and antibodies were purified from antisera. Western blotting was used to identify the protein as marker in two-dimensional electrophoretogram of protoscolex. RESULTS: SDS-PAGE displayed that the proteins separated from Echinococcus granulosus protoscoleces mainly distributed in the M, region of 18,000-90,000. 240 proteins were obtained by two-dimensional electrophoresis with M, 15,790-117,050 and IP 4.0-9.5, and 85.8% (206/241) of the proteins showed the IP ranged from 5 to 9. Western blotting showed that the specific antibody of rEg14-3-3 identified the 14-3-3 protein in two-dimensional electrophoretogram of protoscolex with Mr 33 000 and IP 4.86, the specific antibody of rEgZW-5 identified the ZW-5 protein with Mr 23,000 and IP 4.98, and the specific antibody of rEg P-29 identified the P-29 protein with Mr 29,000 and IP 5.65. CONCLUSION: The antibodies against the three recombinant proteins from Echinococcus granulosus protoscoleces can identify corresponding proteins in the two-dimensional electrophoregrams.