Biosciences, College of Life and Environmental Sciences, University of Exeter, UK.
Analyst. 2020 Apr 7;145(7):2751-2757. doi: 10.1039/c9an02516b. Epub 2020 Feb 24.
A multiplexed biophotonic assay platform has been developed using the localised particle plasmon in gold nanoparticles assembled in an array and functionalised for two assays: total IgG and C-reactive protein (CRP). A protein A/G (PAG) assay, calibrated with a NIST reference material, shows a maximum surface coverage of θ = 7.13 ± 0.19 mRIU, equivalent to 1.5 ng mm of F(ab)-presenting antibody. The CRP capture antibody has an equivalent surface binding density of θ = 2.95 ± 0.41 mRIU indicating a 41% capture antibody availability. Free PAG binding to the functionalised anti-CRP surface shows that only 47 ± 3% of CRP capture antibodies are correctly presenting Fab regions for antigen capture. The accuracy and precision of the CRP sensor assay was assessed with 54 blood samples containing spiked CRP in the range 2-160 mg L. The mean accuracy was 0.42 mg L with Confidence Interval (CI) at 95% from -14.7 to 13.8 mg L and the precision had a Coefficient of Variation (CV) of 10.6% with 95% CI 0.9%-20.2%. These biophotonic platform performance metrics indicate a CRP assay with 2-160 mg L dynamic range, performed in 8 minutes from 5 μL of whole blood without sample preparation.
已开发出一种基于金纳米粒子局部粒子等离子体的多重生物光子分析平台，该平台组装成阵列，并针对两种分析物进行了功能化：总 IgG 和 C 反应蛋白（CRP）。使用 NIST 参考物质校准的蛋白 A/G（PAG）分析显示，最大表面覆盖率θ=7.13±0.19 mRIU，相当于 1.5 ng mm 的 F(ab)呈现抗体。CRP 捕获抗体的等效表面结合密度θ=2.95±0.41 mRIU，表明捕获抗体的可用性为 41%。游离 PAG 与功能化抗 CRP 表面的结合表明，只有 47±3%的 CRP 捕获抗体正确呈现 Fab 区域用于抗原捕获。CRP 传感器分析的准确性和精密度使用含有 2-160 mg L 范围内 CRP 的 54 个血液样本进行评估。平均准确度为 0.42 mg L，置信区间（CI）为 95%，从-14.7 到 13.8 mg L，精密度的变异系数（CV）为 10.6%，95%CI 为 0.9%-20.2%。这些生物光子平台性能指标表明 CRP 分析具有 2-160 mg L 的动态范围，可在 8 分钟内从 5 μL 全血中完成，无需样品制备。
