血清载脂蛋白B椭偏光蛋白质芯片技术定量检测的条件优化

发布时间：2018-04-03 14:10

  本文选题：椭偏光蛋白芯片技术　切入点：光学蛋白质芯片　出处：《山东医药》2016年47期

【摘要】：目的优化椭偏光蛋白芯片技术定量检测血清载脂蛋白B(ApoB)的条件,并观察最佳条件下椭偏光蛋白芯片技术定量检测血清ApoB的准确性。方法用椭偏光蛋白芯片技术检测含有0.2、0.3、0.4、0.5、0.6 mg/m L的载脂蛋白B抗体(anti ApoB)和200μg/m L ApoB标准品的溶液,测得灰度值最高者选为最佳anti-ApoB浓度。用椭偏光蛋白芯片技术检测5、10、20、50、100、200μg/m L ApoB标准品溶液,anti-ApoB浓度为0.3 mg/m L,结果以灰度值表示。然后在上述标准品溶液中加入蛋白G使其终浓度为0.1 mg/m L,再检测一次灰度值。比较两次检测的灰度值,确定是否应该加入0.1 mg/m L蛋白G。最佳条件下椭偏光蛋白芯片技术检测0、5、10、20、50、100、200μg/m L ApoB标准品溶液,计算上述ApoB标准品溶液灰度值与不含ApoB的空白探针溶液(含anti ApoB和蛋白G)灰度值的差值。对上述ApoB标准品溶液浓度和灰度值差值的相关性进行分析,以ApoB标准品溶液浓度为横坐标,灰度值差值为纵坐标描画标准曲线并建立方程。最佳条件下用椭偏光蛋白芯片技术检测16份40倍稀释的已知ApoB浓度(采用免疫比浊法测定)血清的灰度值两次。用上述方程计算16份血清中ApoB浓度,并与其实际ApoB浓度比较。结果最佳anti-ApoB浓度为0.3 mg/m L。与不应用蛋白G者相比,应用0.1 mg/m L蛋白G的椭偏光蛋白芯片技术测得样品的灰度值高(P均0.05)。以ApoB浓度为横坐标x、灰度差值为纵坐标y描画标准曲线,Maple9.5软件得出的回归方程为:y=-23.116Ln(1-x/60),R2=0.936 4。16份血清ApoB浓度和第一次、第二次用椭偏光蛋白芯片技术检测测得血清ApoB浓度相比P均0.05。结论椭偏光蛋白芯片技术检测血清ApoB的最佳条件为anti-ApoB浓度0.3 mg/m L,蛋白G浓度0.1 mg/m L。此条件下椭偏光蛋白芯片技术检测血清ApoB结果准确。
[Abstract]:Objective to optimize the conditions for quantitative detection of serum apolipoprotein (ApoB) by ellipsometry protein chip technique and to observe the accuracy of ellipsometry protein chip technology for quantitative detection of serum ApoB.Methods the solution of Apolipoprotein B antibody anti ApoB and 200 渭 g / mL ApoB standard was detected by ellipsometry protein chip technique. The best anti-ApoB concentration was selected as the highest grayscale value of Apolipoprotein B antibody (ApoB) and 200 渭 g / mL ApoB.The concentration of anti-ApoB in ApoB standard solution was 0.3 mg/m / L using ellipsometry protein chip technique. The results were expressed as gray scale.The final concentration of protein G was 0. 1 mg/m / L, and the gray value was detected again.The gray values of the two tests were compared to determine whether 0.1 mg/m L protein G should be added.Under the optimum conditions, the standard solution of ApoB was detected by ellipsometry protein chip technique. The difference between the gray value of the ApoB standard solution and that of the blank probe solution (including anti ApoB and protein G) without ApoB was calculated.The correlation between the concentration of the ApoB standard solution and the gray value difference was analyzed. The standard curve was drawn and the equation was established with the concentration of the ApoB standard solution as the horizontal coordinate and the gray value difference as the vertical coordinate.Under the optimum conditions, the gray values of 16 serum samples with 40 times dilution of known ApoB concentration (determined by immune turbidimetry) were determined twice by ellipsometry protein chip technique.The ApoB concentration in 16 serum samples was calculated by the above equation and compared with the actual ApoB concentration.Results the optimum concentration of anti-ApoB was 0.3 mg/m L.Compared with those without protein G, the grayscale values of 0. 1 mg/m L protein G were measured by ellipsometry protein chip technique.浠�ApoB娴撳害涓烘í鍧愭爣x,鐏板害宸�澹�

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