A high sensitivity assay for the inflammatory marker C-Reactive protein employing acoustic biosensing |
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Authors: | Jeffrey D McBride Matthew A Cooper |
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Institution: | (1) Akubio Ltd., 181 Cambridge Science Park, Cambridge, CB4 0GJ, UK |
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Abstract: | C-Reactive Protein (CRP) is an acute phase reactant routinely used as a biomarker to assess either infection or inflammatory
processes such as autoimmune diseases. CRP also has demonstrated utility as a predictive marker of future risk of cardiovascular
disease. A new method of immunoassay for the detection of C-Reactive Protein has been developed using Resonant Acoustic Profiling™
(RAP™) with comparable sensitivity to a high sensitivity CRP ELISA (hsCRP) but with considerable time efficiency (12 minutes
turnaround time to result). In one method, standard solutions of CRP (0 to 231 ng/mL) or diluted spiked horse serum sample
are injected through two sensor channels of a RAP™ biosensor. One contains a surface with sheep antibody to CRP, the other
a control surface containing purified Sheep IgG. At the end of a 5-minute injection the initial rate of change in resonant
frequency was proportional to CRP concentration. The initial rates of a second sandwich step of anti-CRP binding were also
proportional to the sample CRP concentration and provided a more sensitive method for quantification of CRP. The lower limit
of detection for the direct assay and the homogenous sandwich assay were both 20 ng/mL whereas for the direct sandwich assay
the lower limit was 3 ng/mL. In a step towards a rapid clinical assay, diluted horse blood spiked with human CRP was passed
over one sensor channel whilst a reference standard solution at the borderline cardiovascular risk level was passed over the
other. A semi-quantities ratio was thus obtained indicative of sample CRP status. Overall, the present study revealed that
CRP concentrations in serum that might be expected in both normal and pathological conditions can be detected in a time-efficient,
label-free immunoassay with RAP™ detection technology with determined CRP concentrations in close agreement with those determined
using a commercially available high sensitivity ELISA. |
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