Preparation And Identification Of Recombinant Antigens Of IgG4 Of Resisting Cysticercus Cellulosae

Cysticercus cellulosae is Zoonoses caused by Cysticercus cellulosae which is larva of Taenia solium.The harm degree of Cysticercus cellulosae varies according to part and amount. Cerebral cysticercosis is of great harm.The effective diagnosis of Cysticercus cellulosae is key to control it.But as a Platyhelminthes parasite,antigenic component of Cysticercus cellulosae is very complex.Cross-reactivity with other helminth infections occurs in wholewormantigen and affects specificity of detection. Cysticercus antigen includes. C.Cyst fluid antigen ,C.Scolex antigen and C.Baldder wall antigen.These antigens vary with different developmental stage . Moreover ,excretary-secretary antigen was excreted during the development of cysticercus in host body . Even so,recent year with the development of molecular hiology,recombinant antigens has become hotspot of immunodiagnosis of cysticercosis because of favorable detection effect. Compared with C.Scolex and C.Baldder , cyst fluid of Cysticercus cellulosae is full of lipoprotein and glycoprotein .PH value of antigenic component is mainly located in basicity region .In recent years,immunologic studies shows that a specific IgG4 level was elevated when the body was stimulated by a complex antigen includingsome parasites.After the recovery from the infection ,this specific IgG4 level fell rapidly or disappeared.This suggests the specific IgG4 level reflected the infectious condition of the patients. Forein scholars have found that in some parasitie infections,specific IgG increases in all four subclasses antibody but especially in IgG4, Over 50% of the total reactive antibody was IgG4,and this was found in almost all patients. In this paper,C.Cyst fluid antigen was used to conduct dimensional electrophoresis and immunoblotting. It is found that alkaline protein bind tightly to gG4 antibody, primarily small molecular weight proein .In the study,51 antigen spots was screened by antigen.24 positive antigen spots were found by mass spectrographic analysis.These spots were related with 12 kinds of protein of cysticercus.Every positive spot has 4 to 5 related protein . Secreted antigen Ts8B3( 16/24 ), Secreted antigen Ts8B2 ( 18/24 ), Sequence 3 from patent US 6589752(15/24)和Cysticercosis 10kDa antigen(16/24)were the most related protein. The ES antigen gene Ts8B2 was cloned from Cysticercus cellulosae by RT-PCR and sequenced. The cDNA fragment without signal peptide was subcloned intoPET30α-GST vector and expressed in E.coli BL-21.SDS-PAGE showed that the expressed fusion protein was approximately 33ku.High purity recombinant protein was obtained by GST affinity chromatography. This study laid foundation for further study of the recombinant protein as antigen for antibody detection of Cysticercus cellulosae.