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Accuracy and precision analysis for a biophotonic assay of C-reactive protein

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Abstract

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.

Graphical abstract: Accuracy and precision analysis for a biophotonic assay of C-reactive protein

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Article information


Submitted
12 Dec 2019
Accepted
17 Feb 2020
First published
18 Feb 2020

Analyst, 2020, Advance Article
Article type
Paper

Accuracy and precision analysis for a biophotonic assay of C-reactive protein

P. James-Pemberton, U. Łapińska, M. Helliwell, R. V. Olkhov, O. J. Hedaux, C. J. Hyde and A. M. Shaw, Analyst, 2020, Advance Article , DOI: 10.1039/C9AN02516B

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