本文選題：AMOZ　切入點：單克隆抗體　出處：《暨南大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：用自制的抗AMOZ單克隆抗體建立檢測組織樣品中AMOZ殘留的間接競爭ELISA和膠體金免疫層析法。 方法：AMOZ與對醛基苯甲酸(4-CBA)反應(yīng),衍生成半抗原CP-AMOZ,采用活化酯法將CPAMOZ與載體蛋白牛血清白蛋白(BSA)、卵清蛋白(OVA)分別偶聯(lián)制備完全抗原,高效液相色譜質(zhì)譜法(LC-MS)、SDS-PAGE、紫外光譜掃描鑒定抗原合成效果。通過免疫BALB/c小鼠、雜交瘤制備技術(shù)制備抗CPAMOZ的單克隆抗體,并對其效價、亞類、特異性等方面進(jìn)行鑒定,建立檢測組織樣品中AMOZ殘留的間接競爭ELISA和膠體金免疫層析檢測方法。 結(jié)果：1.成功合成AMOZ的衍生物CPAMOZ及完全抗原CPAMOZ-BSA及CPAMOZ-OVA。2.成功獲得穩(wěn)定分泌抗CPAMOZ單克隆抗體的雜交瘤細(xì)胞株,該細(xì)胞株分泌的抗體效價高,屬于IgGl類抗體,與CPAMOZ特異性結(jié)合,與載體蛋白BSA、OVA及硝基呋喃類代謝物及其衍生物交叉反應(yīng)均小于1%。3.建立了檢測AMOZ的間接競爭ELISA法,ICso為0.86μg/L,最低檢測限(LOD)為0.07μg/L,組織樣品的樣品回收率為81.48%-85.24%；膠體金免疫層析法的最低檢測限為5μg/L。 結(jié)論：1.成功制備了呋喃它酮代謝物AMOZ的完全抗原及單克隆抗體,該抗體靈敏度高,特異性好。2.成功建立了能用于檢測組織樣品中AMOZ殘留的間接競爭ELISA法和快速、靈敏、穩(wěn)定的膠體金免疫層析法。
[Abstract]:Aim: to establish indirect competitive ELISA and colloidal gold immunochromatographic assay for the detection of AMOZ residues in tissue samples with self-made anti-AMOZ monoclonal antibody. Methods CPAMOZ was conjugated with bovine serum albumin (BSA) and ovalbumin (ovalbumin) to prepare complete antigen by the reaction of: Amoz with 4-CBA), and then the hapten CP-AMOZ was derived into CP-AMOZ by activation ester method, which was coupled with bovine serum albumin (BSA) and ovalbumin (ovalbumin) respectively to prepare complete antigen. High performance liquid chromatography-mass spectrometry (HPLC / MS) / SDS-PAGE and UV scanning were used to identify the effect of antigen synthesis. Monoclonal antibodies against CPAMOZ were prepared by immunizing BALB/c mice and hybridoma preparation technique, and their titers, subclasses and specificity were identified. To establish indirect competitive ELISA and colloidal gold immunochromatography for the detection of AMOZ residues in tissue samples. Results 1. CPAMOZ, a derivative of AMOZ, and its complete antigen CPAMOZ-BSA and CPAMOZ-OVA.2.The hybridoma cell lines secreting monoclonal antibodies against CPAMOZ were successfully obtained. The hybridoma cells secreted high titers of antibodies, which belonged to IgGl class antibodies and specifically bound to CPAMOZ. The cross reaction with the carrier protein BSA-OVA, nitrofuran metabolites and their derivatives was less than 1.3.The indirect competitive ELISA method for the detection of AMOZ was established. The ELISA method was 0.86 渭 g / L, the lowest detection limit was 0.07 渭 g / L, and the sample recovery of tissue samples was 81.48 and 85.24.The colloidal gold immune layer was established. The detection limit of the method is 5 渭 g / L. Conclusion the complete antigen and monoclonal antibody of furantadone metabolite AMOZ were successfully prepared. The antibody was sensitive and specific. 2. An indirect competitive ELISA method was successfully established for the detection of AMOZ residues in tissue samples. Stable colloidal gold immunochromatography.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R392.1

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