Detection of adenosine triphosphate in HeLa cell using capillary electrophoresis-laser induced fluorescence detection based on aptamer and graphene oxide

• A GO-based aptasensor for ATP detection was proposed using CE-LIF detecting technique.
• Compared with using fluorometer, CE-LIF possesses merits of high efficiency, small sample consumption and high sensitivity.
• The method had been successfully applied to quantification of ATP in HeLa cell.

A method for ATP quantification based on dye-labeled aptamer/graphene oxide (aptamer/GO) using capillary electrophoresis-laser induced fluorescence (CE-LIF) detecting technique has been established. In this method, the carboxyfluorescein (FAM)-labelled ATP aptamers were adsorbed onto the surface of GO, leading to the fluorescence quenching of FAM; after the incubation with a limited amount of ATP, stronger affinity between ATP aptamer and ATP resulted in the desorption of aptamers and the fluorescence restoration of FAM. Then, aptamer-ATP complex and excess of aptamer/GO and GO were separated and quantified by CE-LIF detection. It was shown that a linear relation was existing in the CE-LIF peak intensity of aptamer-ATP and ATP concentration in range of 10–700 μM, the regression equation was F = 1.50 + 0.0470C(ATP) (R2 = 0.990), and the limit of detection was 1.28 μM (3S/N, n = 5), which was one order magnitude lower than that of detection in solution by fluorescence method. The approach with excellent specificity and reproducibility has been successfully applied to detecting concentration of ATP in HeLa cell.

Aptamer/GO and CE-LIF were combined to determine ATP, by CE the recovered fluorescence would no longer be quenched by GO.Figure optionsDownload as PowerPoint slide