Detection of adenosine triphosphate in HeLa cell using capillary electrophoresis-laser induced fluorescence detection based on aptamer and graphene oxide
| Autor: | Chao-Yang Wang, Chun-Yuan Wang, Fang Chen, Bi-Yun Fang, Ming-Hao Yao, Yuan-Di Zhao |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Aptamer
Analytical chemistry 02 engineering and technology Microscopy Atomic Force 01 natural sciences chemistry.chemical_compound Adenosine Triphosphate Colloid and Surface Chemistry Capillary electrophoresis Spectroscopy Fourier Transform Infrared Humans Physical and Theoretical Chemistry Fluorescein Laser-induced fluorescence Detection limit Lasers SELEX Aptamer Technique 010401 analytical chemistry Electrophoresis Capillary Reproducibility of Results Oxides Surfaces and Interfaces General Medicine Aptamers Nucleotide Fluoresceins 021001 nanoscience & nanotechnology Fluorescence 0104 chemical sciences Microscopy Fluorescence chemistry Biophysics Graphite 0210 nano-technology Adenosine triphosphate HeLa Cells Biotechnology |
| Zdroj: | Colloids and Surfaces B: Biointerfaces. 140:233-238 |
| ISSN: | 0927-7765 |
| DOI: | 10.1016/j.colsurfb.2015.12.043 |
| Popis: | 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) (R(2)=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. |
| Databáze: | OpenAIRE |
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