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基于散射式生物光子微阵列成像的抗体定量无标记筛选

Quantitative label-free screening for antibodies using scattering biophotonic microarray imaging.

机构信息

School of Biosciences, University of Exeter, Exeter EX4 4QD, UK.

出版信息

Anal Biochem. 2010 Jan 1;396(1):30-5. doi: 10.1016/j.ab.2009.08.008. Epub 2009 Aug 11.

Abstract

A biophotonic array based on gold nanoparticles functionalized with antigen proteins has been used to determine the concentrations of the respective antibodies in solution. Four proteins-fibrinogen, bovine serum albumin, transferrin, and C-reactive protein-were used to construct a test array with the assay repeated a number of times. The antibody-antigen association and dissociation rate constants were determined for the antibody assays from a series of calibration experiments. The label-free determination of the unknown antibody concentrations was performed using two related kinetic analyses. From these results, the current array assay sensitivity is 250 ng ml(-1) with an accuracy of 15% using an 8-min kinetic measurement and a 16-spot averaged assay.

摘要

一种基于金纳米粒子的生物光子阵列,其表面功能化有抗原蛋白,可用于测定溶液中相应抗体的浓度。该阵列使用了四种蛋白质(纤维蛋白原、牛血清白蛋白、转铁蛋白和 C 反应蛋白),通过多次重复实验构建了一个测试阵列。通过一系列校准实验,确定了用于抗体分析的抗体-抗原缔合和解离速率常数。使用两种相关的动力学分析方法对未知抗体浓度进行了无标记测定。根据这些结果,使用 8 分钟的动力学测量和 16 个点的平均测定,当前的阵列分析灵敏度为 250ng/ml,准确率为 15%。

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