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Accuracy and precision analysis for a biophotonic assay of C-reactive protein.
Analyst ( IF 3.6 ) Pub Date : 2020/02/18 , DOI: 10.1039/c9an02516b Philip James-Pemberton 1 , Urszula Łapińska , Mark Helliwell , Rouslan V Olkhov , Oliver J Hedaux , Christopher J Hyde , Andrew M Shaw
Analyst ( IF 3.6 ) Pub Date : 2020/02/18 , DOI: 10.1039/c9an02516b Philip James-Pemberton 1 , Urszula Łapińska , Mark Helliwell , Rouslan V Olkhov , Oliver J Hedaux , Christopher J Hyde , Andrew M Shaw
Affiliation
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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 θmax = 7.13 ± 0.19 mRIU, equivalent to 1.5 ng mm-2 of F(ab)-presenting antibody. The CRP capture antibody has an equivalent surface binding density of θmax = 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-1. The mean accuracy was 0.42 mg L-1 with Confidence Interval (CI) at 95% from -14.7 to 13.8 mg L-1 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-1 dynamic range, performed in 8 minutes from 5 μL of whole blood without sample preparation.
中文翻译：
C反应蛋白生物光子测定的准确性和精密度分析。
已经开发出了一种多重生物光子测定平台，该平台使用阵列中组装的金纳米颗粒中的局部粒子等离激元进行了功能化，可用于两种测定：总IgG和C反应蛋白（CRP）。用NIST参考材料校准的蛋白质A / G（PAG）分析显示最大表面覆盖度为θmax= 7.13±0.19 mRIU，相当于1.5 ng mm-2的F（ab）呈递抗体。CRP捕获抗体的等效表面结合密度为θmax= 2.95±0.41 mRIU，表明捕获抗体的利用率为41％。与功能化的抗CRP表面结合的游离PAG显示，只有47±3％的CRP捕获抗体正确呈现了用于抗原捕获的Fab区。使用包含2-160 mg L-1范围内加标CRP的54个血液样品评估了CRP传感器测定的准确性和精密度。平均准确度为0.42 mg L-1，置信区间（CI）为-14.7至13.8 mg L-1时为95％，精确度的变异系数（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分析的动态范围为2-160 mg L-1，可在5分钟内从5μL全血中进行，无需样品制备。
更新日期：2020-03-31
中文翻译：
C反应蛋白生物光子测定的准确性和精密度分析。
已经开发出了一种多重生物光子测定平台，该平台使用阵列中组装的金纳米颗粒中的局部粒子等离激元进行了功能化，可用于两种测定：总IgG和C反应蛋白（CRP）。用NIST参考材料校准的蛋白质A / G（PAG）分析显示最大表面覆盖度为θmax= 7.13±0.19 mRIU，相当于1.5 ng mm-2的F（ab）呈递抗体。CRP捕获抗体的等效表面结合密度为θmax= 2.95±0.41 mRIU，表明捕获抗体的利用率为41％。与功能化的抗CRP表面结合的游离PAG显示，只有47±3％的CRP捕获抗体正确呈现了用于抗原捕获的Fab区。使用包含2-160 mg L-1范围内加标CRP的54个血液样品评估了CRP传感器测定的准确性和精密度。平均准确度为0.42 mg L-1，置信区间（CI）为-14.7至13.8 mg L-1时为95％，精确度的变异系数（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分析的动态范围为2-160 mg L-1，可在5分钟内从5μL全血中进行，无需样品制备。
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