Development of ImmunoFET Biosensor for Detection of 3-Nitrotyrosine

Khine Kyi Han, Supachoke Mangmool, Korbtham Sathirakul, Kosum Chansiri, Wutthinan Jeamsaksiri, Montri Yasawong

Abstract


3-Nitrotyrosine (3-NT) is one of the important biomarkers for detection of excess of oxidative and nitrosative stress in the body, and derived from the peroxynitrite-induced nitration of free and protein-bound at tyrosine residues. It was increased with oxidative/nitrosative stress-induced apoptosis in various tumor cells and many human diseases. Therefore, the analytical methods to detect 3-NT levels in biological samples are important for in vitro study of the apoptotic mechanism of the compounds and for diagnosis of 3 NT-related diseases. In this study, an accurate, sensitive, and specific assay was developed for 3-NT based on the immunoFET biosensors and the effects of protein-A for orientated immobilization of antibody was investigated on the sensitivity of this biosensors. 
The comparison of sensor performance among two immobilization methods revealed that the assay sensitivity was appreciably improved and better with the antibody attached covalently to the sensor surface than the immobilization method using protein-A. In addition, with this covalent immobilization technique, the output voltage shifts increased linearly (R2 = 0.9923) across the 3-NT concentrations range from 10 to 1000 ng/mL and the limit of quantification of 10 ng/mL and the limit of detection was 0.15 ng/mL. Thus, the developed immunoFET sensors would be applicable for detection of 3-NT in the biological samples.

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