| Sulfonamides(SM) are foremost medicines to control and cure infectious diseases. Sulfadiazine (SD) is representative among sulfonamides. It plays an important role in curing infects, especially influenzal meningitis. SD has a broad antibacterial spectrum and obvious curative effect. The cost being low, SM can be produced without grain.However, accumulative residue of SM in food produce can cause severe problems because of its illegal use, and gives highly rise to concern regarding consumer safety. Normal bacterium will get drug-resistance if people eat contaminated food. Also, it causes intoxication, hypersensitivity. Synthesis of nuclear acid will be disturbed, and people are prone to cancers, especially thyroid carcinoma.Many countries, such as America, European United, Japan and China included, have formulated strict regulations for the use of SM. The residue of SM is a focal point in exports and imports quarantine. An effective and quick detect technology is required to fulfill the quarantine system; to ensure regular exports and imports trades. Also, it will restrict the illegal use of SM, insure food safety, and improve people healthy.High Performance Liquid Chromatography (HPLC) is major method to detect the residue of SM now. HPLC is a simple, convenient and precise technology. However, the pretreatment of samples is complicated and expatiatory. Its cost is quite high. All those have restrict its extensive use. Immune analysis is a new technology based on antigen-antibody special combination, which is highly sensitive, selective, quick and convenient. The immune analysis replies to microanalysis from complicated samples.Monoclonal antibodies are produced by cell fusion technology. In contrast to polyclonal antibodies, monoclonal antibodies have following virtues: identical structures, uniform component, high specificity, animate biotic-activity and fewer cross reacts.Monoclonal antibodies of SD are prospective in this study, with the purpose of providing capital materials for the immune analysis system.Heterogeneity, large molecule colloid, and degradation are necessary conditions of complete antigens. Molecular weight being 250, SD is a half antigen. There is an aromatic amido in molecular of SD, so diazotization is available. Bovine serum albumin (BSA) has complicated constructer, multiple available site, satisfactory antigenicity, strong resistance, well dissolvability, and easy to get. All these reveal BSA an optimal carrier. Ovalbumin is chosed in the same time for controlled trial and detection.Conjugating SD to BSA by diazotization transformed SD, an half antigen, into complete antigen. The production is BSA-SD. Diazotization means that aromatic molecule reacts with nitrite salt at 0~5C, then conjugate to hydroxyl of protein by azo-bond.Adequately dialysis is required to purify the product. Then, UV-spectrum technology is used for confirm that SD has been conjugated to BSA. The foregoing react has altered the constructer of BSA, which has changed its ultraviolet absorption and shape. Its color has changed from achromatous into typical yellow.To gain purified special mice anti-SD, six Balb/C mice were immunized against BSA-SD. Antibody titers were determined using indirect ELISA with OVA-SD in coating the micro-wells.The specific anti-SD sera were detected in all mice and were sufficient for succedent experiment.SP2/0-Ag14 myelomas cell were chosen for cell fusion. Keep it in the bloom of proliferation before cell fusion.Feeder cells should be prepared before cell fusion. Kill one or more Balb/C mouse and draw its celiac macrophage and lymphocyte, then pave them in the cell culture wells. These should be operated one day before cell fusion.Electro-fusion, laser-fusion, chem.-fusion and bio-fusion are optional. Chem.-fusion is chosen. Highly purified Polyethylene 1000(PEG-1000) was used for cell fusion.Cell fusion was proceeded in a 50-ml centrifuge tube. Mingle spleen cells and myeloma cells (5:1) into the centrifuge tube, and then instill PEG-1000 scrupulously at a certain...
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