|
|
|
|
|
|
A New Label-Free Fluorometric Assay for ATP Based on Split Aptamer |
LI Fei-fei1,3, LU Yi-song2, YANG Sheng-yuan1,3*, LIN Xi1,3, CHEN Wei1, LIU Can1,3, XIAO Fu-bing1,3, LIANG Hao1,3 |
1. College of Public Health,University of South China,Hengyang 421001,China
2. Affiliated Nanhua Hospital, University of South China, Hengyang 421001, China
3. Key Laboratory of Hengyang for Health Hazard Factors Inspection and Quarantine, Hengyang 421001,China |
|
|
Abstract A novel Label-Free Fluorometric Assay based on the recombination of split aptamer chip was developed for thedetection of adenosine triphosphate (ATP). In this strategy, the split aptamer was selected as a specific capture probe for the split two fragments aptamers could specifically form a ternary assembly in the presence of ligand and the two separate oligonucleotides lack secondary structures, thus not yielding false-positive or nonspecific signals, while the Thiazole orange(TO), an almost non-fluorescence dye in buffer solution, was used as signal probe, and the single-walled carbon nanotubes (SWCNTs) was applied to reduce the background signals. In the pH 8.0 Tris-HCl buffer solution, those two split aptamer fragments will be combined with each other to form a stable “aptamer-ATP-aptamer” composite structure upon interacting with its target ATP. The “sandwich” structure can’t wrap the sidewalls of the SWCNTs and is freed in solution, and TO shows agreat fluorescence enhancement when binding to the “aptamer-ATP-aptamer” composite structure. In the absence of ATP, the split aptamers, existing in a single-stranded state, bind to the surface of the SWCNTs via a π—π-conjugate interaction, and TO shows weak fluorescence because “sandwich”structure is not formed. In the system, the higher the ATP concentration is, the more the “aptamer-ATP-aptamer” sandwich recognition structure complex obtained, sois the fluorescence. Under the optimized experimental conditions, the ATP concentration in the range from 9.0×10-9 mol·L-1 to 1.0×10-7 mol·L-1 was linear with the ΔF/F0 value at the maximum fluorescence emission wavelength of 550 nm, r=0.996 4, with a low detection limit of 2.67×10-9 mol·L-1. The recoveries of the method were 95.2%~104%, and the relative standard deviation (RSD) was 1.02%~4.54%, respectively. Based on the specific molecular recognition and high affinity of twosplit aptamers, the reaction product was shown that a “turn-on” fluorescence response to ATP with good selectivity, only a slight fluorescence change could be observed by GTP, CTP, and UTP (at a 200-fold higher concentration than that of ATP), indicating that UTP, CTP, and GTP could not interact with P1 and P1 to initiate the reaction. The method is simple, rapid, free-label, sensitive and accurate, and can be used for the determination of ATP in serum samples. Therefore, the present strategy has a great potential application prospect in the field of rapid detection of small molecular substances.
|
Received: 2018-08-02
Accepted: 2018-12-12
|
|
Corresponding Authors:
YANG Sheng-yuan
E-mail: yangshyhy@126.com
|
|
[1] Wang G,Su X,Xu Q,et al. Biosensors & Bioelectronics,2018,101:129.
[2] Ding H,Xiong Y,Sun J,et al. Frontiers in Neuroscience, 2018, 12: 431.
[3] Varik V, Oliveira S R A, Hauryliuk V. Scientific Reports, 2017, 7: 11022.
[4] Zhou L,Gan N,Wu Y,et al. Analyst,2018,143(11):2696.
[5] Ma C H,Lin C S,Wang Y R,et al. Trends in Analytical Chemistry,2016,77: 226.
[6] Huizenga D E. Biochemistry,1995,34(2):656.
[7] Duan W,Wang X,Wang H. Talanta,2018,180:76.
[8] Xu L, Shen X, Li B, et al. Analytica Chimica Acta, 2017, 980: 58.
[9] Wang H,Chen H,Huang Z,et al. Talanta,2018,184:219.
[10] WU Xi,PEI Xiao-jing,LIN Ruo-yun,et al(吴 熙,裴晓静,林若韵,等). Spectroscopy and Spectral Analysis(光谱学与光谱分析),2017,37(1):13.
[11] Bagheri H,Afkhami A,Khoshsafar H,et al. Biosensors& Bioelectronics,2017,89(2): 829.
[12] Li Q,Wang Y D,Shen G L,et al. Chemical Communications (Cambridge,England),2015,51(20):4196.
[13] Yuan B,Zhou Y,Guo Q,et al. Chemical Communications (Cambridge, England),2016,52(8):1590.
[14] Chen A L,Yan M M,Yang S M. Trends in Analytical Chemistry,2016,80:581.
[15] WANG Qing,DAI Jian-feng,LI Wei-xue,et al(王 青,戴剑锋,李维学,等). Journal of Lanzhou University of Technology(兰州理工大学学报),2006,(3):157. |
[1] |
LEI Hong-jun1, YANG Guang1, PAN Hong-wei1*, WANG Yi-fei1, YI Jun2, WANG Ke-ke2, WANG Guo-hao2, TONG Wen-bin1, SHI Li-li1. Influence of Hydrochemical Ions on Three-Dimensional Fluorescence
Spectrum of Dissolved Organic Matter in the Water Environment
and the Proposed Classification Pretreatment Method[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2024, 44(01): 134-140. |
[2] |
XIA Ming-ming1, 2, LIU Jia3, WU Meng1, 2, FAN Jian-bo1, 2, LIU Xiao-li1, 2, CHEN Ling1, 2, MA Xin-ling1, 2, LI Zhong-pei1, 2, LIU Ming1, 2*. Three Dimensional Fluorescence Characteristics of Soluble Organic Matter From Different Straw Decomposition Products Treated With Calcium Containing Additives[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2024, 44(01): 118-124. |
[3] |
GU Yi-lu1, 2,PEI Jing-cheng1, 2*,ZHANG Yu-hui1, 2,YIN Xi-yan1, 2,YU Min-da1, 2, LAI Xiao-jing1, 2. Gemological and Spectral Characterization of Yellowish Green Apatite From Mexico[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2024, 44(01): 181-187. |
[4] |
HAN Xue1, 2, LIU Hai1, 2, LIU Jia-wei3, WU Ming-kai1, 2*. Rapid Identification of Inorganic Elements in Understory Soils in
Different Regions of Guizhou Province by X-Ray
Fluorescence Spectrometry[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2024, 44(01): 225-229. |
[5] |
LIU Wei1, 2, ZHANG Peng-yu1, 2, WU Na1, 2. The Spectroscopic Analysis of Corrosion Products on Gold-Painted Copper-Based Bodhisattva (Guanyin) in Half Lotus Position From National Museum of China[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3832-3839. |
[6] |
WANG Hong-jian1, YU Hai-ye1, GAO Shan-yun1, LI Jin-quan1, LIU Guo-hong1, YU Yue1, LI Xiao-kai1, ZHANG Lei1, ZHANG Xin1, LU Ri-feng2, SUI Yuan-yuan1*. A Model for Predicting Early Spot Disease of Maize Based on Fluorescence Spectral Analysis[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3710-3718. |
[7] |
CHENG Hui-zhu1, 2, YANG Wan-qi1, 2, LI Fu-sheng1, 2*, MA Qian1, 2, ZHAO Yan-chun1, 2. Genetic Algorithm Optimized BP Neural Network for Quantitative
Analysis of Soil Heavy Metals in XRF[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3742-3746. |
[8] |
SONG Yi-ming1, 2, SHEN Jian1, 2, LIU Chuan-yang1, 2, XIONG Qiu-ran1, 2, CHENG Cheng1, 2, CHAI Yi-di2, WANG Shi-feng2,WU Jing1, 2*. Fluorescence Quantum Yield and Fluorescence Lifetime of Indole, 3-Methylindole and L-Tryptophan[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3758-3762. |
[9] |
WANG Zhi-qiang1, CHENG Yan-xin1, ZHANG Rui-ting1, MA Lin1, GAO Peng1, LIN Ke1, 2*. Rapid Detection and Analysis of Chinese Liquor Quality by Raman
Spectroscopy Combined With Fluorescence Background[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3770-3774. |
[10] |
YI Min-na1, 2, 3, CAO Hui-min1, 2, 3*, LI Shuang-na-si1, 2, 3, ZHANG Zhu-shan-ying1, 2, 3, ZHU Chun-nan1, 2, 3. A Novel Dual Emission Carbon Point Ratio Fluorescent Probe for Rapid Detection of Lead Ions[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3788-3793. |
[11] |
YANG Ke-li1, 2, PENG Jiao-yu1, 2, DONG Ya-ping1, 2*, LIU Xin1, 2, LI Wu1, 3, LIU Hai-ning1, 3. Spectroscopic Characterization of Dissolved Organic Matter Isolated From Solar Pond[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3775-3780. |
[12] |
QI Guo-min1, TONG Shi-qian1, LIN Xu-cong1, 2*. Specific Identification of Microcystin-LR by Aptamer-Functionalized Magnetic Nanoprobe With Laser-Induced Fluorescence[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3813-3819. |
[13] |
HE Yan-ping, WANG Xin, LI Hao-yang, LI Dong, CHEN Jin-quan, XU Jian-hua*. Room Temperature Synthesis of Polychromatic Tunable Luminescent Carbon Dots and Its Application in Sensitive Detection of Hemoglobin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(11): 3365-3371. |
[14] |
LIN Hong-jian1, ZHAI Juan1*, LAI Wan-chang1, ZENG Chen-hao1, 2, ZHAO Zi-qi1, SHI Jie1, ZHOU Jin-ge1. Determination of Mn, Co, Ni in Ternary Cathode Materials With
Homologous Correction EDXRF Analysis[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(11): 3436-3444. |
[15] |
LI Xiao-li1, WANG Yi-min2*, DENG Sai-wen2, WANG Yi-ya2, LI Song2, BAI Jin-feng1. Application of X-Ray Fluorescence Spectrometry in Geological and
Mineral Analysis for 60 Years[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(10): 2989-2998. |
|
|
|
|