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    摘要：通过用特异性抗氯噻啉的单克隆抗体淘选噬菌体展示肽库获得751种噬菌体展示肽。选择克隆L4-29建立噬菌体酶免疫吸附分析（P-ELISA）和噬菌体化学发光酶免疫分析（P-CLEIA）。 P-ELISA和P-CLEIA的最佳缓冲液为pH 8，0.14 M NaCl和2.5％甲醇的PBS缓冲液。P-ELISA的半数最大抑制浓度（IC50）值，检测限（LOD，IC10值）和线性范围（IC10-IC90）分别为1.45,0.55和0.55至3.82 ng mL -1，P-CLEIA 分别为0.86,0.13和0.13-5.84 ng mL -1。与传统ELISA相比，P-ELISA和P-CLEIA的灵敏度分别提高了40倍和68倍。P-CLEIA比P-ELISA更敏感，具有更宽的线性范围。两种分析方法与氯噻啉的类似物（除吡虫啉）没有显著的交叉反应性。P-ELISA和P-CLEIA两种方法在稻田水，土壤，甘蓝，水稻，苹果，梨和番茄样品中对氯噻啉的添加回收率分别为72.3-101.3％和73.9-102.6％。P-ELISA和P-CLEIA与高效液相色谱（HPLC）对样品中氯噻啉检出结果的相关性良好，可用于实际样品中的氯噻啉的检测。34621
    毕业论文关键词：氯噻啉;酶免疫吸附分析;化学发光酶免疫分析;噬菌体展示肽;模拟表位
    Development of phage enzyme immunoassays for the determination of imidaclothiz
    Abstract：Six phage-displayed peptides were obtained by biopanning the phage-displayed peptide libraries with the specific anti-imidaclothiz monoclonal antibody. A phage enzyme-linked immunosorbent assay (P-ELISA) and a phage chemiluminescent enzyme immunoassay (P-CLEIA) for imidaclothiz were developed by using a clone L4-29. The optimal buffers of P-ELISA and P-CLEIA were pH 8, PBS buffer containing 0.14 M NaCl and 2.5% methanol. The half maximal inhibition concentration (IC50) values, limit of detection (LOD, IC10 values) and linear range (IC10 to IC90) of the P-ELISA were 1.45, 0.55 and 0.55 to 3.82 ng mL-1, while the P-CLEIA were 0.86, 0.13 and 0.13 to 5.84 ng mL-1, respectively. Compared with conventional ELISAs, the sensitivities of P-ELISA and P-CLEIA were increased by more than 40-fold and 68-fold, respectively. The P-CLEIA was more sensitive and showed wider linear range than the P-ELISA. Two analyses had no significant cross-reactivity with the analogues of imidaclothiz except for imidacloprid. Recoveries of the P-ELISA and P-CLEIA for imidaclothiz in paddy water, soil, cabbage, rice, apple, pakchoi, pear and tomato samples were 72.3-101.3% and 73.9-102.6%, respectively. The P-ELISA and P-CLEIA were feasible to detect the amounts of imidaclothiz in the authentic samples, which showed good correlation with that obtained from high-performance liquid chromatography (HPLC).
    Keywords：Imidaclothiz, Enzyme-linked immunosorbent assay, Chemiluminescent enzyme immunoassay, Phage-displayed peptide, Peptidomimetics
    目录
    摘要    2
    关键词    2
    Abstract    2
    Keywords    2
    引言    2
    1.材料与方法     4
    1.1材料     4
    1.2主要仪器    4
    1.3方法     4
    1.3.1氯噻啉单克隆抗体的制备     4
    1.3.2生物淘选     4
    1.3.3噬菌体酶免疫吸附分析方法     5
    1.3.4噬菌体化学发光酶免疫分析方法     5
    1.3.5P-ELISA和P-CLEIA的优化     5
    1.3.6标准曲线的建立     5
    1.3.7交叉反应性（CR）     5
    1.3.8 加标样品的分析     5
    1.3.9 HPLC分析验证    6
    2.结果与分析     6
    2.1噬菌体展示肽的淘选     6
    2.2噬菌体展示肽的选择     6
    2.3 P-ELISA和P-CLEIA的优化    7
    2.4 P-ELISA和P-CLEIA的灵敏度     7
    2.5 P-ELISA和P-CLEIA的特异性     8
    2.6加标样品的分析     9
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