Preparation And Biological Characteristics Identification Of Toxoplasma Gondii Monoclonal Antibodies

Toxoplasma gondii is a global zoonotic parasitic diseases.The prevalence and circulation of this parasitic disease not only influence the development of stock raising severely, but also menace the health of human beings.The purpose of this experiment is to prepare monoclonal antibodies against Toxoplasma gondii by mainly using the protein expression and hybridoma technique in immunology and molecular biology.This work would set up the foundation for further study on establishing diagnosis and prevention and cure of Toxoplasmosis.Recombinant plasmid pET32a-P30was Constructed by PCR, cloning, sequencing of the P30gene.Then the prokaryotic expression of recombinant plasmid pET32a-P30was successful induced and Toxoplasma gondii P30recombinant protein antigen was purified.Also Toxoplasma gondii tachyzoite antigen was made by trypsin method.Albino rats were immuned by using P30recombinant protein antigen and tachyzoite antigen of Toxoplasma gondii. Four hybridoma cell lines, named as 5B6,2D25D5and2A1of Toxoplasma gondii parasite antigen were respectively obtained by hybridoma technique and an indirect ELISA detecting method.The biological characteristics of four hybridoma cell lines were identified: The cell culture supernatant titer of5B6,2D2,5D5,2A1are1:800,1:1600,1:3200,1:6400and the ascites antibody titer of5B6.2D2,5D5,2Alare1:104,1:104,1:105,1:105repectitvely by indirect ELISA detecting method.The results showed the number of chromosome of the hybridoma cell lines were from90to110. The prepared four McAbs were found to be of the isotypes of IgG1, IgM, Ig G1and IgM repectitvely.By indirect ELISA detection and SPSS statistical software for analysis it is proved that the first and the tenth generations of four hybridoma cell lines can secret monoclonal antibody stablelyGuangxi Zhuang Autonomous Region is the epidemic areas of humans and animals toxoplasmosis. In this study, we had prepared P30recombinant protein antigen and tachyzoite antigen of Toxoplasma gondii.By hybridoma technique and using these antigen we had prepared monoclonal antibody of Toxoplasma gondiiand its biological characteristics were identified.Our work had laid the foundation for establishing rapid and sensitive detection methods and immune prophylaxis of toxoplasmosis.