A rapid detection of neopterin based on a label-free and homogeneous FRET immunoassay system

Herein, we have developed a label-free and homogeneous fluorescence resonance energy transfer (FRET) immunoassay for the detection of neopterin (NPT), which is an early and valuable biochemical marker of cellular immunity. Owing to intrinsic fluorescence properties of antibody and NPT, anti-NPT antibody (anti-NPT) and analyte played roles as the respective donor and acceptor in the FRET immunoassay. As the concentration of NPT increases, the fluorescence intensity at ~350 nm decreases owing to the formation of increasing amounts of the anti-NPT/NPT complex in which FRET takes place. The assay system was found to display a high specificity and a low detection limit (0.14 ng mL-1) for NPT. A practical application of the FRET immunoassay system was demonstrated by its use in the detection of NPT in spiked human serum samples. The observations made in these efforts show that the homogeneous FRET immunoassay strategy, which requires a simple sample preparation procedure, serves as a powerful tool for the rapid and sensitive quantitative determination of NPT.

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