蛋白芯片检测牛乳β-乳球蛋白条件的优化

发布时间：2018-03-25 03:26

本文选题：蛋白芯片　切入点：β-乳球蛋白　出处：《卫生研究》2017年01期

【摘要】：目的优化蛋白芯片检测β-乳球蛋白(β-Lg)的技术条件。方法用芯片点样仪将β-乳球蛋白的一种抗体点样于三维基片上,用其另一种抗体作为检测抗体,以Cy3标记的羊抗作为二抗进行检测。以双抗夹心法对β-Lg抗原进行检测。结果β-Lg单克隆鼠抗66#被确定为点样探针,选择接触式点样进行芯片点样,点样数在42~92点之间可出现较好的点样一致性;β-Lg固定探针的浓度选择0.5 mg/mL,其检测抗体的稀释度为1∶2000;1%无蛋白封闭液被优选为封闭剂;β-Lg检测下限与生物检测限分别为17.54和55.31 ng/mL。根据β-Lg的S型曲线确定了其线性范围并建立了对β-Lg具有最佳决定系数(R~2=0.9993)的回归方程与标准曲线。结论本研究优化了牛乳中β-Lg检测的蛋白芯片检测条件,建立了定量检测牛乳β-Lg的蛋白芯片平台。
[Abstract]:Objective to optimize the technical conditions for the detection of 尾 -lactoglobulin (尾 -Lg) by using a protein chip. The Cy3 labeled goat antibody was used as the second antibody, and the 尾 -Lg antigen was detected by double antibody sandwich method. The concentration of 尾 -Lg fixed probe was 0.5 mg / mL, the dilution of antibody was 1: 2000 1% and the detection limit of 尾 -Lg was 17.54, while the concentration of 尾 -Lg fixed probe was 17.54, and the concentration of 尾 -Lg fixed probe was 0.5 mg / mL, and the dilution of antibody was 1: 2000 1%, the concentration limit of 尾 -Lg was 17.54, and the concentration of 尾 -Lg was 17.54, and the concentration of 尾 -Lg was 17.54, respectively. And 55.31 ng / mL. According to the S-shape curve of 尾 -Lg, the linear range was determined and the regression equation and standard curve of 尾 -Lg were established. Conclusion the detection conditions of 尾 -Lg in milk by protein chip were optimized. A protein chip platform for quantitative detection of 尾-LG in milk was established.
【作者单位】：中国疾病预防控制中心营养与健康所;
【基金】：中国疾病预防控制中心青年科研基金(No.2015A202)
【分类号】：R155.5

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