Optimization of ellipsometry protein-chip technique in quantitative detection of serum apolipoprotein B
NING Xiang
WANG Jiaojiao
WANG Juan
BI Shaojie
SHENG Lin
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.
Keywords：ellipsometry protein-chip techniqueoptical protein-chipellipsometry imagingApoBProtein GQuantitative detection
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 17-20 )