Optimization of Aptamer-Based Electrochemical Biosensor for ATP Detection Using Screen-Printed Carbon Electrode/Gold Nanoparticles (SPCE/AuNP)

Rahmaniar Mulyani; Nida Yumna; Iman Permana Maksum; Toto Subroto; Yeni Wahyuni Hartati

doi:10.22146/ijc.72820

Optimization of Aptamer-Based Electrochemical Biosensor for ATP Detection Using Screen-Printed Carbon Electrode/Gold Nanoparticles (SPCE/AuNP)

https://doi.org/10.22146/ijc.72820

Rahmaniar Mulyani⁽¹⁾, Nida Yumna⁽²⁾, Iman Permana Maksum⁽³⁾, Toto Subroto⁽⁴⁾, Yeni Wahyuni Hartati^(5*)

(1) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang Km. 21, Jatinangor 45363, West Java, Indonesia; Department of Chemistry, Faculty of Sciences and Informatics, Universitas Jenderal Achmad Yani, Jl. Terusan Jenderal Sudirman, Cimahi 40531, West Java, Indonesia
(2) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang Km. 21, Jatinangor 45363, West Java, Indonesia
(3) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang Km. 21, Jatinangor 45363, West Java, Indonesia
(4) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang Km. 21, Jatinangor 45363, West Java, Indonesia
(5) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang Km. 21, Jatinangor 45363, West Java, Indonesia
(*) Corresponding Author

Abstract

Electrochemical biosensors are used to detect adenosine triphosphate (ATP) levels, which are involved in a variety of biological processes, such as regulating cellular metabolism and biochemical pathways. Therefore, this research aims to develop an aptamer-based electrochemical biosensor with Screen Printed Carbon Electrode/gold nanoparticles (SPCE/AuNP) and collect data as well as information related to ATP detection. The modification of SPCE with AuNP increased the analyte’s binding sensitivity and biocompatibility. The aptamer was selected based on its excellent bioreceptor characteristics. Furthermore, aptamer–SH (F1) and aptamer-NH₂ (F2) were immobilized on the SPCE/AuNP surface, which had been characterized using SEM, EIS, and DPV. Also, the ATP-binding aptamers were electrochemically characterized using the K₃[Fe(CN)₆] redox system and Differential Pulse Voltammetry (DPV). According to the optimization results using the Box-Behnken experimental design, the ideal conditions obtained from the factors influencing the experiment were the F1 concentration and incubation time of 4 µM and 24 h, respectively, as well as F1/F2/ATP incubation time of 7.5 min. Meanwhile, for the range of 0.1 to 100 µM, the detection (LoD) and quantification (LoQ) limits were 7.43 and 24.78 µM, respectively. Therefore, this aptasensor method can be used to measure ATP levels in real samples.

Keywords

adenosine triphosphate (ATP); aptamer; AuNP; electrochemistry; screen printed carbon electrode (SPCE)

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References

[1] Huang, Y., Lei, J., Cheng, Y., and Ju, H., 2015, Target-assistant Zn²⁺-dependent DNAzyme for signal-on electrochemiluminescent biosensing, Electrochim. Acta, 155, 341–347.

[2] Frazier, A.E., Thorburn, D.R., and Compton, A.G., 2019, Mitochondrial energy generation disorders: Genes, mechanisms, and clues to pathology, J. Biol. Chem., 294 (14), 5386–5395.

[3] Maksum, I.P., Farhani, A., Rachman, S.D., and Ngili, Y., 2013, Making of the A3243G mutant template through site directed mutagenesis as positive control in PASA-Mismatch three bases, Int. J. PharmTech Res., 5 (2), 441–450.

[4] Maksum, I.P., Natradisastra, G., Nuswantara, S., and Ngili, Y., 2013, The effect of A3243G mutation of mitochondrial DNA to the clinical features of type-2 diabetes mellitus and cataract, Eur. J. Sci. Res., 96 (4), 591–599.

[5] Hartati, Y.W., Nur Topkaya, S., Maksum, I.P., and Ozsoz, M., 2013, Sensitive detection of mitochondrial DNA A3243G tRNALeu mutation via an electrochemical biosensor using Meldola’s Blue as a hybridization indicator, Adv. Anal. Chem., 3 (A), 20–27.

[6] Destiarani, W., Mulyani, R., Yusuf, M., and Maksum, I.P., 2020, Molecular dynamics simulation of T10609C and C10676G mutations of mitochondrial ND4L gene associated with proton translocation in type 2 diabetes mellitus and cataract patients, Bioinf. Biol. Insights, 14, 117793222097867.

[7] Maksum, I.P., Saputra, S.R., Indrayati, N., Yusuf, M., and Subroto, T., 2017, Bioinformatics study of m.9053G>A mutation at the ATP6 gene in relation to type 2 diabetes mellitus and cataract diseases, Bioinf. Biol. Insights, 11, 1177932217728515.

[8] Ning, Y., Wei, K., Cheng, L., Hu, J., and Xiang, Q., 2017, Fluorometric aptamer based determination of adenosine triphosphate based on deoxyribonuclease I-aided target recycling and signal amplification using graphene oxide as a quencher, Microchim. Acta, 184 (6), 1847–1854.

[9] Qu, F., Sun, C., Lv, X., and You, J., 2018, A terbium-based metal-organic framework@gold nanoparticle system as a fluorometric probe for aptamer based determination of adenosine triphosphate, Microchim. Acta, 185 (8), 359.

[10] Liu, X., Lin, B., Yu, Y., Cao, Y., and Guo, M., 2018, A multifunctional probe based on the use of labeled aptamer and magnetic nanoparticles for fluorometric determination of adenosine 5’-triphosphate, Microchim. Acta, 185 (4), 243.

[11] Khlyntseva, S.V., Bazel’, Y.R., Vishnikin, A.B., and Andruch, V., 2009, Methods for the determination of adenosine triphosphate and other adenine nucleotides, J. Anal. Chem., 64 (7), 657–673.

[12] Huang, Y.F., and Chang, H.T., 2007, Analysis of adenosine triphosphate and glutathione through gold nanoparticles assisted laser desorption/ionization mass spectrometry, Anal. Chem., 79 (13), 4852–4859.

[13] Srivastava, P., Razi, S.S., Ali, R., Srivastav, S., Patnaik, S., Srikrishna, S., and Misra, A., 2015, Highly sensitive cell imaging “Off–On” fluorescent probe for mitochondria and ATP, Biosens. Bioelectron., 69, 179–185.

[14] Chen, J.R., Jiao, X.X., Luo, H.Q., and Li, N.B., 2013, Probe-label-free electrochemical aptasensor based on methylene blue-anchored graphene oxide amplification, J. Mater. Chem. B, 1 (6), 861–864.

[15] Yi, Q., and Yu, W., 2009, Nanoporous gold particles modified titanium electrode for hydrazine oxidation, J. Electroanal. Chem., 633 (1), 159–164.

[16] Yáñez-Sedeño, P., Villalonga, R., and Pingarrón, J.M., 2015, "Electroanalytical Methods Based on Hybrid Nanomaterials" in Encyclopedia of Analytical Chemistry, John Wiley & Sons, Ltd, Chichester, UK, 1–18.

[17] Goud, K.Y., Moonla, C., Mishra, R.K., Yu, C., Narayan, R., Litvan, I., and Wang, J., 2019, Wearable electrochemical microneedle sensor for continuous monitoring of levodopa: Toward Parkinson management, ACS Sens., 4 (8), 2196–2204.

[18] Villalonga, A., Pérez-Calabuig, A.M., and Villalonga, R., 2020, Electrochemical biosensors based on nucleic acid aptamers, Anal. Bioanal. Chem., 412 (1), 55–72.

[19] Mashhadizadeh, M.H., Naseri, N., and Mehrgardi, M.A., 2017, A simple non-enzymatic strategy for adenosine triphosphate electrochemical aptasensor using silver nanoparticle-decorated graphene oxide, J. Iran. Chem. Soc., 14 (9), 2007–2016.

[20] Zheng, J., Li, X., Wang, K., Song, J., and Qi, H., 2020, Electrochemical nanoaptasensor for continuous monitoring of ATP fluctuation at subcellular level, Anal. Chem., 92 (16), 10940–10945.

[21] Metters, J.P., Kadara, R.O., and Banks, C.E., 2011, New directions in screen printed electroanalytical sensors: An overview of recent developments, Analyst, 136 (6), 1067.

[22] Mulyasuryani, A., and Dofir, M., 2014, Enzyme biosensor for detection of organophosphate pesticide residues base on screen printed carbon electrode (SPCE)-bovine serum albumin (BSA), Engineering, 6 (5), 230–235.

[23] Taleat, Z., Khoshroo, A., and Mazloum-Ardakani, M., 2014, Screen-printed electrodes for biosensing: A review (2008–2013), Microchim. Acta, 181 (9), 865–891.

[24] Dorledo de Faria, R.A., Messaddeq, Y., Heneine, G.D., and Matencio, T., 2019, Application of screen-printed carbon electrode as an electrochemical transducer in biosensors, Int. J. Biosens. Bioelectron., 5 (1), 1–2.

[25] Kanyong, P., Rawlinson, S., and Davis, J., 2016, Gold nanoparticle modified screen-printed carbon arrays for the simultaneous electrochemical analysis of lead and copper in tap water, Microchim. Acta, 183 (8), 2361–2368.

[26] Dridi, F., Marrakchi, M., Gargouri, M., Saulnier, J., Jaffrezic-Renault, N., and Lagarde, F., 2017, "Nanomaterial-Based Electrochemical Biosensors for Food Safety and Quality Assessment" in Nanobiosensors, Eds. Grumezescu, A.M., Academic Press, Cambridge, US, 167–204.

[27] Zhao, X., Mai, Z., Kang, X., and Zou, X., 2008, Direct electrochemistry and electrocatalysis of horseradish peroxidase based on clay–chitosan-gold nanoparticle nanocomposite, Biosens. Bioelectron., 23 (7), 1032–1038.

[28] Bernardo-Boongaling, V.R.R., Serrano, N., García-Guzmán, J.J., Palacios-Santander, J.M., and Díaz-Cruz, J.M., 2019, Screen-printed electrodes modified with green-synthesized gold nanoparticles for the electrochemical determination of aminothiols, J. Electroanal. Chem., 847, 113184.

[29] Kashefi-Kheyrabadi, L., and Mehrgardi, M.A., 2013, Aptamer-based electrochemical biosensor for detection of adenosine triphosphate using a nanoporous gold platform, Bioelectrochemistry, 94, 47–52.

[30] Miller, J.N., 1991, Basic statistical methods for analytical chemistry. Part 2. Calibration and regression methods, A review, Analyst, 116 (1), 3–14.

[31] Skoog, D.A., Holler, F.J., and Nieman, T.A., 1998, Principles of Instrumental Analysis, 5^th Ed., Saunders College Publishing, Philadelphia, US.

[32] Hwu, S., Garzuel, M., Forró, C., Ihle, S.J., Reichmuth, A.M., Kurdzesau, F., and Vörös, J., 2020, An analytical method to control the surface density and stability of DNA-gold nanoparticles for an optimized biosensor, Colloids Surf., B, 187, 110650.

[33] Zhao, P., Li, N., and Astruc, D., 2013, State of the art in gold nanoparticle synthesis, Coord. Chem. Rev., 257 (3-4), 638–665.

[34] Apyari, V.V., Arkhipova, V.V., Dmitrienko, S.G., and Zolotov, Y.A., 2014, Using gold nanoparticles in spectrophotometry, J. Anal. Chem., 69 (1), 1–11.

[35] Fatimah, S., Haryati, I., and Jamaludin, A., 2009, Pengaruh Uranium terhadap Analisis Thorium menggunakan Spektrofotometer UV-Vis, Seminar Nasional V SDM Teknologi Nuklir, 5 November 2009, Yogyakarta, 573–578.

[36] Yáñez-Sedeño, P., and Pingarrón, J.M., 2005, Gold nanoparticle-based electrochemical biosensors, Anal. Bioanal. Chem., 382 (4), 884–886.

[37] Ren, L., Xu, P., Zhang, P., Qin, Z., Zhang, Y., and Jiang, L., 2021, Label-free fluorescence aptasensor based on AuNPs and CQDs for the detection of ATP, AIP Adv., 11, 015316.

[38] Shahsavar, K., Hosseini, M., Shokri, E., Ganjali, M.R., and Ju, H., 2017, A sensitive colorimetric aptasensor with a triple-helix molecular switch based on peroxidase-like activity of a DNAzyme for ATP detection, Anal. Methods, 9 (32), 4726–4731.

DOI: https://doi.org/10.22146/ijc.72820