毕业论文化学发光免疫分析.doc

q 河北农业大学 本 科 毕 业 论 文 题 目： 化学发光免疫分析用于抗人白蛋白单体的克隆筛选 学 院： 生命科学学院 专业班级： 制药工程1001 学 号： 2010044030111 学生姓名： 石冰卓 指导教师姓名： 张冬冬 指导教师职称： 讲师 二〇一四年六月一日 化学发光免疫分析用于抗人白蛋白单抗的克隆筛选 制药工程1001班：石冰卓 指导老师：摘要：本实验将化学发光免疫分析用于小鼠抗人白蛋白单克隆抗体（anti-ALB McAb）的克隆筛选。方法简化为一步法操作，通过正交实验[L9（34）表]优化实验参数，寻找阳性克隆/空白对照发光值（信噪比）最大时的参数组合，并对该组合进行了方法学评价及统计学分析。结果表明包被抗体为3mg/L, 酶标蛋白A稀释比例为1：2000，温育时间30分钟时信噪比最大为1284，数倍高于其它组合。方法学评价表明该实验条件下检测灵敏度为20ng/L；其线性范围为 20-20000ng/L；精密度良好，平均批内CV为5.32%，平均批间CV为8.82%。实验结果证实该方法可用于anti-ALB McAb的克隆筛选，且方法学上明显优于ELISA等传统方法，特别是一步操作的简便快速，也适合其它抗体高通量筛选。 关键词：化学发光免疫分析、抗人白蛋白单克隆抗体、正交试验、克隆筛选Chemiluminescence immune assay?for?screening and cloning of?anti human albumin?Monoclonal Antibody Abstract: the chemiluminescent immunoassay for monoclonal antibody of mouse anti human albumin (anti-ALB McAb) cloning and screening. Simplified method for?one-step operation,?through the orthogonal?experiment [L9?(34)?surface] optimization of experimental parameters,?find the?positive clone/blank control the light value(SNR) parameters of maximum and the combination of analysis and statistical methodology.?The results?showed that the?coating antibody?to 3mg/L, the enzyme labeled?protein A?dilution?ratio was 1:2000,?the incubation time?of 30minutes when the?signal-to-noise ratio?is?1284,?is several times higher than that of?other combination.?The evaluation?shows that the?detection?sensitivity of the experimental conditions?for the 20ng/L method;?the linear?range is 20-20000ng/L; good precision,?the average?number of?CV?was 5.32%,?the average?number of CV?is 8.82%.?The experiment results prove that this method?can be used for screening?and cloning of the anti-ALB?McAb,?and the?method?is obviously better than the ELISA?and other traditional methods,?especially the?step of operation?is simple and fast,?but also for?other?antibody?screening. Keywords:?chemiluminescence immune?analysis; screening of anti?human albumin?monoclonal an