[Purification of recombinant Schistosoma japonicum signaling protein 14-3-3 and antibody preparation].
OBJECTIVE
To prepare and characterize the polyclonal and monoclonal antibodies(McAbs) against Schistosoma japonicum(Sj) signaling protein 14-3-3.


METHODS
Recombinant Sj14-3-3 protein was expressed and the gel slices were lyophilized. Rabbits were immunized with the ground gel powder to prepare polyclonal antibodies against Sj14-3-3. BALB/c mice were immunized with recombinant Sj14-3-3 purified by electroelution. The anti-rSj14-3-3 monoclonal antibody was obtained by using hybridoma technique. Characterization of the antibodies was performed by enzyme linked immunosorbent assay (ELISA) and Western blotting analysis.


RESULTS
Purified recombinant Sj14-3-3 protein was obtained. The titers of immune sera from the animals were 1:8-1:64. The monoclonal antibody(McAb) against 14-3-3 antigen was obtained and its subclass was found to be IgG1. The McAb reacted strongly and specifically with the expressed Sj14-3-3 protein.


CONCLUSIONS
The anti-14-3-3 sera were obtained and the hybridoma cell lines which secreted stable monoclonal antibodies against Sj14-3-3 were established, which provides a new approach to study the role of Sj14-3-3 in the signal transduction of S. japonicum.
Li
Feng
F
Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan 430060.
Hu
Min
M
Shen
Ji-long
JL
D048948:14-3-3 Proteins; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D005260:Female; D006825:Hybridomas / Q000378:metabolism; D007106:Immune Sera; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification*; D012549:Schistosoma japonicum / Q000276:immunology*; D014446:Tyrosine 3-Monooxygenase / Q000276:immunology / Q000302:isolation & purification*
D048948:14-3-3 Proteins; D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D014446:Tyrosine 3-Monooxygenase