Abstract
We present an immunoassay for Interferon-γ (IFN-γ) with a limit of detection of 1.9 pM (30 pg/mL) and a linear concentration range spanning three orders of magnitude. The developed one-step assay takes only 12 min and can replace the time-consuming and labor-intensive enzyme-linked immunosorbent assay (ELISA). The solid-phase sandwich assay is performed on a new measurement system comprising single-use test tubes and a compact fluorescence reader. The polymer tubes contain an optical configuration for the detection of supercritical angle fluorescence, allowing for highly sensitive real-time binding measurements.
Highlights
Interferon-γ (IFN-γ) is a small, homodimeric protein mainly produced by T-cells and natural killer cells
The calibration curve shown in Figure 3(B) was obtained by plotting the recombinant mouse IFN-γ (rmIFN-γ) concentrations from 10 pM to 5 nM versus the supercritical angle fluorescence (SAF) intensity measured after 700 s
We have developed a rapid and sensitive assay for IFN-γ on the SAF immunoassay platform
Summary
Interferon-γ (IFN-γ) is a small, homodimeric protein mainly produced by T-cells and natural killer cells. It plays key functions in host defense against pathogens by exerting anti-viral, anti-proliferative and immunoregulatory activities. Its sensitive and accurate quantification is relevant in medical research and diagnostics. The most widely used method for the quantification of bioanalytes is ELISA. A typical ELISA protocol involves dozens of washing and incubation steps, takes several hours and requires relatively large amounts of expensive antibody conjugates. The need for better solutions is driving the development of new assay technologies using magnetic micro- and nano-particle [1,2,3,4,5]
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