Preparation Of Synthesis Of Antigen And Monoclonal Antibody On Heavy Metal Ion Cadmium

With the continuous improvement of people's living standards and enhance the environmental awareness, the heavy metal pollution was particularly concerned about and the residue testing should be strengthened. However, the present heavy metal detection method did commonly rely on the large equipments, the trouble pre-treatment, high costs, long time. In this study, the immune antigen and the detective antigen of Cd2+ were synthesized with chelators and carrier protein. The synthesized antigen was accessed by UV spectrophotometry, trinitrobenzene sulfonic acid, and anti-serum titer testing. Cadmium ions ELISA was established by preparation of monoclonal antibody technology. The aim was to provide a convenient, accurate and specific, cheap and fast new technique on heavy metal Cd2+ residues detection.RESULTS: (1) The different absorption peak was observed among chelator, protein and synthesized antigen. The content of free amino was from 33% to 40%. (2) The titer of anti-serum was not increased with dose of antigen. The titer of anti-serum at dose of 100μg per mouse was 1: 81 920, and it was higher than other doses. (3)The best condition of fusion included in a proportion of 1:7.1 between SP2/0 and spleen cells and 50 % PEG. Under this condition, the fusion rate was 98.4%. (4) Only 2 wells (1A1 and 4E7) were positive, when the cell supernatant was detected by BSA-IEDTA-Cd and BSA-IEDTA. The masculine rate was only 0.35%. After been cloned 3 times with 100% masculine rate, the monoclonal antibody of anti-Cd²⁺ were excreted by a strain hybridoma cell 1A1. (5) With detecting of the indirect competitive ELISA, the cells supernatant titer of 1A1 was over 1:1 024 and that of the ascites was over 1:256 000. (6)The protein concentration of ascites was 15.04 mg/mL. The monoclonal antibody was IgG1.CONCLUAION: (1) IEDTA can be uesed for synthesis of immune antigen and detecting antigen on the heavy metal ion Cd²⁺. (2) Carrier KLH was more effective than BSA for immunization. Carrier BSA was more effective than KLH for detection. (3) Reasonable immunization programmes was helpful to generate anti-serum. The positive relation was not observed between the immune effect and the immunization dose. (4) Although the monoclonal antibody was successfully produced, the masculine rate was low and the cloning process was enough stable. Antigen synthesis process still need to be further optimized.