title: Detection of adenosine triphosphate in HeLa cell using capillary electrophoresis-laser induced fluorescence detection based on aptamer and graphene oxide.
abstract: 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.
authors.affiliation: Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
authors.forename: Bi-Yun
authors.initials: BY
authors.lastname: Fang
authors.affiliation: Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
authors.forename: Ming-Hao
authors.initials: MH
authors.lastname: Yao
authors.affiliation: Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
authors.forename: Chun-Yuan
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
authors.forename: Chao-Yang
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China; Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China. Electronic address: zydi@mail.hust.edu.cn.
authors.forename: Yuan-Di
authors.initials: YD
authors.lastname: Zhao
authors.affiliation: School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, Wuhan 430074, PR China. Electronic address: fchen@mail.hust.edu.cn.
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis* / Q000737:chemistry; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D006108:Graphite / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D007834:Lasers; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D010087:Oxides / Q000737:chemistry*; D015203:Reproducibility of Results; D052156:SELEX Aptamer Technique / Q000379:methods; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D052157:Aptamers, Nucleotide; D005452:Fluoresceins; D010087:Oxides; C024098:6-carboxyfluorescein; D006108:Graphite; D000255:Adenosine Triphosphate
keywords: Adenosine triphosphate; Aptamer; Capillary electrophoresis; Graphene oxide