山东医药2016，Vol.56Issue(47)：17-20,4.DOI:10.3969/j.issn.1002-266X.2016.47.005
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
Optimization of ellipsometry protein-chip technique in quantitative detection of serum apolipoprotein B
摘要
Abstract
Objective To optimize the conditions for quantitatively detecting human serum apolipoprotein B via ellip-sometry protein-chip technique and to evaluate the accuracy of this technique under optimum conditions.Methods In our study,we detected the gray values of solutions containing 0.2,0.3,0.4,0.5 and 0.6 mg/mL anti-ApoB and 200 μg/mL ApoB with ellipsometry protein-chip technique,and the optimum concentration was the one with the highest gray value.We detected the gray values of solutions containing 0.3 mg/mL anti-ApoB and 5,10,20,50,100,200 μg/mL ApoB before and after protein G was added in those solutions until its final concentration was 0.1 mg/mL.By comparing the grey val-ues,we determined whether protein G should be added or not.We detected the grey values of solutions containing 0,5, 10,20,50,100,200 μg/mL ApoB via ellipsometry protein-chip technique under optimum conditions and calculated the difference between them and the gray values of the solutions without ApoB (consisted of anti-ApoB and protein G).Then we analyzed the correlations between the concentrations and the differences by drawing the curve and establishing the equa-tion of it with concentration as X-axis and gray value as Y-axis.At last,we detected the gray values of 16 serum samples diluted 40 times twice through ellipsometry protein-chip technique and immunoturbidimetry under the optimum conditions. Then we calculated the concentrations of ApoB in the samples with the equation and compared them with its practical con-centration.Results The optimum concentration of anti-ApoB was 0.3 mg/mL.The gray values in protein G group were higher than that of the contrast group (all P<0.05).The regression equation was y=-23.116Ln(1 -x/60),R2 =0.936 4 by Maple 9.5 software according to the curve.No significantly statistical difference was found between the ApoB concentra-tions detected via ellipsometry protein-chip technique and 16 cases of serum apoB concernttations (all P>0.05).Conclu-sions The optimal conditions for detection of ApoB in serum under ellipsometry protein-chip technique are anti-ApoB with the concentration of 0.3 mg/mL and protein G with the concentration of 0.1 mg/mL.The accuracy of the technique under such conditions is reliable.关键词
椭偏光蛋白芯片技术/光学蛋白质芯片/椭偏光生物显微成像技术/载脂蛋白B/载脂蛋白B抗体/蛋白GKey words
ellipsometry protein-chip technique/optical protein-chip/ellipsometry imaging/ApoB/Protein G/Quantitative detection分类
医药卫生引用本文复制引用
宁翔,王娇娇,王娟,毕少杰,盛林..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[J].山东医药,2016,56(47):17-20,4.基金项目
国家自然科学基金资助项目（81500232）；山东省医药卫生科技发展计划项目（2014WS0155）；山东省自然科学基金－联合专项资助项目（2014ZRE27456）。 （）
