The Monoclonal Antibodies Preparation And Application On The Related Antigen Protein Of Trichinella Spiralis And Live Cancer H7402

Trichinella spiralis is a parasite of Trichinellidae which infects both people and livestock.According to the existing research materials, Trichinella spiralis has some interference andsuppression effect on several kinds of tumor cells by reducing the proliferation or inducing theapoptosis of tumor cells. Liver cancer can’t be ignored for its high morbidity and mortality, and itwas recognized a threat to the health of the world. However, the vaccine and traditional medicinefor liver cancer didn’t work well enough. For this, getting some biologicals which have highefficiency, low toxicity and less side effects becomes to be people’s urgent need to fight againstliver cancer. Now, researches have shown that there were cross antigens between Trichinellaspiralis and H7402liver cancer cells；we got gene CK-1of Trichinella spiralis by searching thecDNA expression library,and the monoclonal antibody of CK-1shown that it could suppress thegrowth of cancer cells in/out of mice. This showed that there mightbe anti-cancer substances in orbeing excreted by the worms. Therefore, this research aimed to search the cross antigens betweenthe Trichinella spiralis and the H7402liver cancer cells by screening the cDNA expression libraryand observe the inhibiting effect of tumor cell’s growth, which could serve the biological treatmentof liver cancer and the curative effect of Trichinella spiralis against tumors in the future.Based on the method of screening the cDNA expression library of Trichinella spiralis, we got6genes in relation to H7402liver cancer cells, they were cysteine string protein （Tsp₀6120）,golgi-associated plant pathogeneis-related protein1（Tsp₀7885）, seleno-protein T （Tsp₀3801）,DNA excision repair protein ERCC-1（Tsp₀1641）, deoxynucleotidyltransferase interacting protein1（Tsp₀1476） and nucleoredoxin （Tsp₀4425）. After biological analysis, gene seleno-protein Twas choose to start the prokaryotic expression and purification. In the experiment of makingmonoclonal antibody, we got3monoclonal antibodies,including4F1,4H2and4D4. By the meansof the detection of karyotype, types of antibodies and titre analysis,the result showed that thehybridoma chromosome of4H2was104, the antibody titer determinationwe was higher than1:105,and the antibody subtype was IgG2b. We chose4H2to treat cancer during the followingexperiment, such as the MTT and the animal model test. The result showed that the inhibition ratioreach26.7%when the monoclonal antibody got to50μg/100μL in the MTT; at the same time,the inhibition ratio reach80.3%when the monoclonal antibody got to100μg/100μL in theresearch of animal model.