| Immunoassay is based on the specificity of reversability association reaction between antigen and antibody, it have higher sensitivity and specificity, bigger sample flux, faster detection, lower cost, it is generally used in medicine analysis poison analysis and environmental pollutant detection. Antibody is the most important agent in immunoassay, it had considerable effect in the sensitive and specificness of the immune analyses. Monoclone antibody was produced by single inbred cell line, it had specific immune reaction only with one determinant group of the antigen, it had pure character, higher valency and better specificity, it could avoidance the cross reaction in Serology, the detectability was nanogram or even picogram. Mostly medicine poison and environmental pollutant is haptin, it's molecular mass was smaller than 1000, they had reactionogenicity,but no immunogenicity, they kan combined with specific antibody, and formed hapten-antibody complex, coupled the haptin with carrier which had big molecular mass, to got an artificial immunogen, then immunization the animal, preparation the hybridoma cell and it's specificity monoclonal antibody, this kind of monoclonal antibody can be used in many research , such as medical science, immunology, biology and so on, it had great value and potentiality in analytical technique research.Swainsonine was the main constit poisonous of locoweed, it was a hapten, it had small molecular weight (173), specific antibody could be produced by use the immunogen which was linked with big molecular weight carrier by chemical synthesis to induce animal. These made something become true which were deterring of SW, prevention and cure of locoweed poisoning by immunology. However, monoclonal antibody is most effective tool of circulating antigen deterring presently. In the test Balb/C mice were immunized by SW-HSA for the preparation of spleen cell when antibody titer was up to desirable level. Hybridoma cells that can secrete specific antibody of SW were prepared by monoclonal antibody technique, and then monoclonal antibody of SW was prepared, that establish the foundation for immunodetection on SW, prevention and cure of animal toxic disease.1. Balb/c mice were immunized by SW-HSA emulsified with adjuvant. And then mice kept high antibody titer were obtained by indirect ELISA that deter SW antibody. There display different intensity of immunogenicity of SW-HSA. As the method of indirect ELISA, antibody titers in sera of mice were 27, 26, 26, 26 after the third injection, and that of mice were 29, 28, 28, 28 after the fourth injection.2. This test made 3 times cell fusion used SP2/0 and mice spleen cell, the first fusion test had higher fusion ratio,36.7 %3. Antibody titer of cell culture supernatant of which were clonal was assayed by ELISA, and then filting masculine cells. Cell masculine rate of the first confluence were higher, 7.095%.4 One hybridoma cell named 1C3 was obtained after screen, clone culture and 3 times monoclone culture,it had stabilize growth and can excrete a great quantity of SW monoclonal antibody, Valences of monoclonal antibody of supernatant of hybridoma cell 1D5 were detected. That of supernatant was 26. after high passage and 5 times refrigeration and resuscitation, hybridoma cell 1C3 had stable ability of secrete antibody.5 Nuclear type of hybridoma cell was analyzed by colchicines repression method, and the average number of chromosome of 1C3 stain cell was between 45 and 50 couples which approach the sum of chromosome number of two parent cell, these identify that 1C3 stain cell was a crossbred of both parent cells after cell fusion.6. Balb/c mice were given hybridoma cell 1C3 by intraperitoneal injection, and the SW antibody titer in peritoneal fluid was 211.7 Peritoneal fluid (3mL) was purified by ammonium sulfate precipitation method to gain monoclonal antibody, and then protein which obtained was obtained by freeze drying. The purifided result was detected with sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE). It is proved that purified antibody had been obtained. Molecular wieght is about 55 ku and 26 ku, the same as H-chain and L-chain of antibody.8 Relative affinity of monoclonal antibody in supernatant of hybridoma cell 1C3 was detected by indirect ELISA. It was 56μg/mL.
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