[Purification and immunogenicity identification of the recombinant protein related with specific IgE against Schistosoma japonicum].
OBJECTIVE
To purify the specific IgE antibody-related recombinant protein of Schistosoma japonicum and to identify its immunogenicity.


METHODS
The recombinant plasmid Sj43B/pGEX-6p-1 was expressed in E. coli BL 21. The inclusion body of the fusion protein was washed by TNMFX buffer and separated by FPLC. After renaturation, the fusion protein was used to vaccinate the mice. The specific IgG and IgE antibodies were detected by dot-ELISA and Western blotting analysis, respectively.


RESULTS
Most of the proteins mixed with the inclusion body of the recombinant protein could be eliminated by washing with TNMFX buffer. The purified recombinant fusion protein could be obtained by FPLC separation. The experiment on mice immunized with the fusion protein showed that the specific IgE antibody was generated against the target part of the fusion protein, but not the specific IgG antibody.


CONCLUSIONS
The fusion protein expressed by the recombinant plasmid Sj43B/pGEX-6p-1 could induce specific IgE response of the immunized mice.
Wang
Yong
Y
Laboratory of Molecular Immunoparasitology, Institute of Medical Molecular Biology, Nanjing Medical University, Nanjing 210029.
Su
Chuan
C
Zhang
Zhao-song
ZS
Hu
Xue-mei
XM
Liu
Feng
F
Li
Chun-lin
CL
Ji
Min-jun
MJ
Zhu
Xiang
X
Wu
Guan-ling
GL
D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis*; D015801:Helminth Proteins / Q000276:immunology* / Q000302:isolation & purification; D007073:Immunoglobulin E / Q000096:biosynthesis*; D007074:Immunoglobulin G / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000302:isolation & purification; D012549:Schistosoma japonicum / Q000276:immunology*; D014611:Vaccination
D000909:Antibodies, Helminth; D015801:Helminth Proteins; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E