Development of a surface plasmon resonance immunosensor and ELISA for 3-nitrotyrosine in human urine

Abstract

3-Nitrotyrosine (3-NT) is thought to be a relevant biomarker of nitrosative stress which is associated with many inflammatory and chronic diseases. It is necessary to develop confidential method for specific and sensitive 3-NT detection. In this paper, on the basis of anti-3-NT specific antibody, we developed a label-free indirect competitive surface plasmon resonance (SPR) immunosensor and ELISA for the detection of 3-NT. Under the optimized conditions, the SPR immunosensor can obtain a linear range of 0.17-6.07 μg/mL and a limit of detection (LOD) of 0.12 μg/mL while the ELISA can reach 0.33-9.94 μg/mL and a LOD of 0.24 μg/mL.The selectivity of 3-NT was also testified by six kinds of amino acid analogues. Besides, the developed SPR immunosensor was compared thoroughly with a conventional ELISA in spiked analysis of urine samples. Good recoveries and correlation between these two methods were observed (R² = 0.964). Therefore, it is concluded that the automated SPR platform can be applied to quantify 3-NT in biological samples with its sensitivity, accuracy, and real-timing.

Keywords: 3-nitrotyrosine; ELISA; Human urine; Surface plasmon resonance (SPR).