Label-Free Pyrophosphate Recognition with Functionalized Asymmetric Nanopores

[EN] The label¿free detection of pyrophosphate (PPi) anions with a nanofluidic sensing device based on asymmetric nanopores is demonstrated. The pore surface is functionalized with zinc complexes based on two di(2¿picolyl)amine [bis(DPA)] moieties using carbodiimide coupling chemistry. The complexation of zinc (Zn2+) ion is achieved by exposing the modified pore to a solution of zinc chloride to form bis(Zn2+¿DPA) complexes. The chemical functionalization is demonstrated by recording the changes in the observed current¿voltage (I¿V) curves before and after pore modification. The bis(Zn2+¿DPA) complexes on the pore walls serve as recognition sites for pyrophosphate anion. The experimental results show that the proposed nanofluidic sensor has the ability to sense picomolar concentrations of PPi anion in the surrounding environment. On the contrary, it does not respond to other phosphate anions, including monohydrogen phosphate, dihydrogen phosphate, adenosine monophosphate, adenosine diphosphate, and adenosine triphosphate. The experimental results are described theoretically by using a model based on the Poisson¿Nernst¿Planck equations.M.A., S.N., and W.E. acknowledge the funding from the Hessen State Ministry of Higher Education, Research and the Arts, Germany, under the LOEWE project iNAPO. P.R. and S.M. acknowledge financial support by the Generalitat Valenciana (Program of Excellence Prometeo/GV/0069), the Spanish Ministry of Economic Affairs and Competitiveness (MAT2015-65011-P), and FEDER. I. A. and C.M.N. acknowledge financial support through the Helmholtz programme BioInterfaces in Technology and Medicine. The authors are also thankful to Prof. C. Trautmann, Department of Materials Research from GSI, for support with irradiation experiments.Ali, M.; Ahmed, I.; Ramirez Hoyos, P.; Nasir, S.; Niemeyer, CM.; Mafe, S.; Ensinger, W. (2016). Label-Free Pyrophosphate Recognition with Functionalized Asymmetric Nanopores. Small. 12(15):2014-2021. https://doi.org/10.1002/smll.201600160201420211215Gyurcsányi, R. E. (2008). Chemically-modified nanopores for sensing. TrAC Trends in Analytical Chemistry, 27(7), 627-639. doi:10.1016/j.trac.2008.06.002Hou, X., Guo, W., & Jiang, L. (2011). Biomimetic smart nanopores and nanochannels. Chemical Society Reviews, 40(5), 2385. doi:10.1039/c0cs00053aHou, X., & Jiang, L. (2009). Learning from Nature: Building Bio-Inspired Smart Nanochannels. ACS Nano, 3(11), 3339-3342. doi:10.1021/nn901402bHou, X., Zhang, H., & Jiang, L. (2012). Building Bio-Inspired Artificial Functional Nanochannels: From Symmetric to Asymmetric Modification. Angewandte Chemie International Edition, 51(22), 5296-5307. doi:10.1002/anie.201104904Ali, M., Neumann, R., & Ensinger, W. (2010). Sequence-Specific Recognition of DNA Oligomer Using Peptide Nucleic Acid (PNA)-Modified Synthetic Ion Channels: PNA/DNA Hybridization in Nanoconfined Environment. ACS Nano, 4(12), 7267-7274. doi:10.1021/nn102119qAli, M., Schiedt, B., Neumann, R., & Ensinger, W. (2010). Biosensing with Functionalized Single Asymmetric Polymer Nanochannels. Macromolecular Bioscience, 10(1), 28-32. doi:10.1002/mabi.200900198Ali, M., Yameen, B., Neumann, R., Ensinger, W., Knoll, W., & Azzaroni, O. (2008). Biosensing and Supramolecular Bioconjugation in Single Conical Polymer Nanochannels. Facile Incorporation of Biorecognition Elements into Nanoconfined Geometries. Journal of the American Chemical Society, 130(48), 16351-16357. doi:10.1021/ja8071258Fologea, D., Gershow, M., Ledden, B., McNabb, D. S., Golovchenko, J. A., & Li, J. (2005). Detecting Single Stranded DNA with a Solid State Nanopore. Nano Letters, 5(10), 1905-1909. doi:10.1021/nl051199mIqbal, S. M., Akin, D., & Bashir, R. (2007). Solid-state nanopore channels with DNA selectivity. Nature Nanotechnology, 2(4), 243-248. doi:10.1038/nnano.2007.78Mara, A., Siwy, Z., Trautmann, C., Wan, J., & Kamme, F. (2004). An Asymmetric Polymer Nanopore for Single Molecule Detection. Nano Letters, 4(3), 497-501. doi:10.1021/nl035141oStorm, A. J., Storm, C., Chen, J., Zandbergen, H., Joanny, J.-F., & Dekker, C. (2005). Fast DNA Translocation through a Solid-State Nanopore. Nano Letters, 5(7), 1193-1197. doi:10.1021/nl048030dVlassiouk, I., Kozel, T. R., & Siwy, Z. S. (2009). Biosensing with Nanofluidic Diodes. Journal of the American Chemical Society, 131(23), 8211-8220. doi:10.1021/ja901120fBayley, H., Braha, O., Cheley, S., & Gu, L.-Q. (2005). Engineered Nanopores. Nanobiotechnology, 93-112. doi:10.1002/3527602453.ch7Bayley, H., Braha, O., & Gu, L.-Q. (2000). Stochastic Sensing with Protein Pores. Advanced Materials, 12(2), 139-142. doi:10.1002/(sici)1521-4095(200001)12:23.0.co;2-qBayley, H., & Cremer, P. S. (2001). Stochastic sensors inspired by biology. Nature, 413(6852), 226-230. doi:10.1038/35093038Dekker, C. (2007). Solid-state nanopores. Nature Nanotechnology, 2(4), 209-215. doi:10.1038/nnano.2007.27Healy, K., Schiedt, B., & Morrison, A. P. (2007). Solid-state nanopore technologies for nanopore-based DNA analysis. Nanomedicine, 2(6), 875-897. doi:10.2217/17435889.2.6.875Siwy, Z. S., & Howorka, S. (2010). Engineered voltage-responsive nanopores. Chem. Soc. Rev., 39(3), 1115-1132. doi:10.1039/b909105jSpohr, R. (2005). Status of ion track technology—Prospects of single tracks. Radiation Measurements, 40(2-6), 191-202. doi:10.1016/j.radmeas.2005.03.008Korchev, Y. E., Bashford, C. L., Alder, G. M., Apel, P. Y., Edmonds, D. T., Lev, A. A., … Pasternak, C. A. (1997). A novel explanation for fluctuations of ion current through narrow pores. The FASEB Journal, 11(7), 600-608. doi:10.1096/fasebj.11.7.9212084Siwy, Z., Apel, P., Dobrev, D., Neumann, R., Spohr, R., Trautmann, C., & Voss, K. (2003). Ion transport through asymmetric nanopores prepared by ion track etching. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, 208, 143-148. doi:10.1016/s0168-583x(03)00884-xSiwy, Z. S. (2006). Ion-Current Rectification in Nanopores and Nanotubes with Broken Symmetry. Advanced Functional Materials, 16(6), 735-746. doi:10.1002/adfm.200500471Apel, P. Y., Korchev, Y. ., Siwy, Z., Spohr, R., & Yoshida, M. (2001). Diode-like single-ion track membrane prepared by electro-stopping. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, 184(3), 337-346. doi:10.1016/s0168-583x(01)00722-4Siwy, Z., Trofin, L., Kohli, P., Baker, L. A., Trautmann, C., & Martin, C. R. (2005). Protein Biosensors Based on Biofunctionalized Conical Gold Nanotubes. Journal of the American Chemical Society, 127(14), 5000-5001. doi:10.1021/ja043910fHan, C., Hou, X., Zhang, H., Guo, W., Li, H., & Jiang, L. (2011). Enantioselective Recognition in Biomimetic Single Artificial Nanochannels. Journal of the American Chemical Society, 133(20), 7644-7647. doi:10.1021/ja2004939Liu, Q., Xiao, K., Wen, L., Lu, H., Liu, Y., Kong, X.-Y., … Jiang, L. (2015). Engineered Ionic Gates for Ion Conduction Based on Sodium and Potassium Activated Nanochannels. Journal of the American Chemical Society, 137(37), 11976-11983. doi:10.1021/jacs.5b04911Han, C., Su, H., Sun, Z., Wen, L., Tian, D., Xu, K., … Jiang, L. (2013). Biomimetic Ion Nanochannels as a Highly Selective Sequential Sensor for Zinc Ions Followed by Phosphate Anions. Chemistry - A European Journal, 19(28), 9388-9395. doi:10.1002/chem.201300200Pérez-Mitta, G., Albesa, A. G., Knoll, W., Trautmann, C., Toimil-Molares, M. E., & Azzaroni, O. (2015). Host–guest supramolecular chemistry in solid-state nanopores: potassium-driven modulation of ionic transport in nanofluidic diodes. Nanoscale, 7(38), 15594-15598. doi:10.1039/c5nr04645aTian, Y., Hou, X., Wen, L., Guo, W., Song, Y., Sun, H., … Zhu, D. (2010). A biomimetic zinc activated ion channel. Chemical Communications, 46(10), 1682. doi:10.1039/b918006kShang, Y., Zhang, Y., Li, P., Lai, J., Kong, X.-Y., Liu, W., … Jiang, L. (2015). DNAzyme tunable lead(ii) gating based on ion-track etched conical nanochannels. Chemical Communications, 51(27), 5979-5981. doi:10.1039/c5cc00288eLiu, Q., Xiao, K., Wen, L., Dong, Y., Xie, G., Zhang, Z., … Jiang, L. (2014). A Fluoride-Driven Ionic Gate Based on a 4-Aminophenylboronic Acid-Functionalized Asymmetric Single Nanochannel. ACS Nano, 8(12), 12292-12299. doi:10.1021/nn506257cXie, G., Xiao, K., Zhang, Z., Kong, X.-Y., Liu, Q., Li, P., … Jiang, L. (2015). A Bioinspired Switchable and Tunable Carbonate-Activated Nanofluidic Diode Based on a Single Nanochannel. Angewandte Chemie International Edition, 54(46), 13664-13668. doi:10.1002/anie.201505269Ngo, H. T., Liu, X., & Jolliffe, K. A. (2012). Anion recognition and sensing with Zn(ii)–dipicolylamine complexes. Chemical Society Reviews, 41(14), 4928. doi:10.1039/c2cs35087dLee, S., Yuen, K. K. Y., Jolliffe, K. A., & Yoon, J. (2015). Fluorescent and colorimetric chemosensors for pyrophosphate. Chemical Society Reviews, 44(7), 1749-1762. doi:10.1039/c4cs00353eKim, S. K., Lee, D. H., Hong, J.-I., & Yoon, J. (2009). Chemosensors for Pyrophosphate. Accounts of Chemical Research, 42(1), 23-31. doi:10.1021/ar800003fHargrove, A. E., Nieto, S., Zhang, T., Sessler, J. L., & Anslyn, E. V. (2011). Artificial Receptors for the Recognition of Phosphorylated Molecules. Chemical Reviews, 111(11), 6603-6782. doi:10.1021/cr100242sHeinonen, J. K. (2001). Biological Role of Inorganic Pyrophosphate. doi:10.1007/978-1-4615-1433-6Timms, A. E. (2002). Genetic studies of disorders of calcium crystal deposition. Rheumatology, 41(7), 725-729. doi:10.1093/rheumatology/41.7.725Doherty, M., Belcher, C., Regan, M., Jones, A., & Ledingham, J. (1996). Association between synovial fluid levels of inorganic pyrophosphate and short term radiographic outcome of knee osteoarthritis. Annals of the Rheumatic Diseases, 55(7), 432-436. doi:10.1136/ard.55.7.432Lomashvili, K. A., Khawandi, W., & O’Neill, W. C. (2005). Reduced Plasma Pyrophosphate Levels in Hemodialysis Patients. Journal of the American Society of Nephrology, 16(8), 2495-2500. doi:10.1681/asn.2004080694Ronaghi, M., Karamohamed, S., Pettersson, B., Uhlén, M., & Nyrén, P. (1996). Real-Time DNA Sequencing Using Detection of Pyrophosphate Release. Analytical Biochemistry, 242(1), 84-89. doi:10.1006/abio.1996.0432Yang, S., Feng, G., & Williams, N. H. (2012). Highly selective colorimetric sensing pyrophosphate in water by a NBD-phenoxo-bridged dinuclear Zn(ii) complex. Organic & Biomolecular Chemistry, 10(29), 5606. doi:10.1039/c2ob25617gLiu, D. J., Credo, G. M., Su, X., Wu, K., Lim, H. C., Elibol, O. H., … Varma, M. (2011). Surface immobilizable chelator for label-free electrical detection of pyrophosphate. Chemical Communications, 47(29), 8310. doi:10.1039/c1cc12073eCredo, G. M., Su, X., Wu, K., Elibol, O. H., Liu, D. J., Reddy, B., … Varma, M. (2012). Label-free electrical detection of pyrophosphate generated from DNA polymerase reactions on field-effect devices. The Analyst, 137(6), 1351. doi:10.1039/c2an15930aAli, M., Bayer, V., Schiedt, B., Neumann, R., & Ensinger, W. (2008). Fabrication and functionalization of single asymmetric nanochannels for electrostatic/hydrophobic association of protein molecules. Nanotechnology, 19(48), 485711. doi:10.1088/0957-4484/19/48/485711Xue, L., Wang, H.-H., Wang, X.-J., & Jiang, H. (2008). Modulating Affinities of Di-2-picolylamine (DPA)-Substituted Quinoline Sensors for Zinc Ions by Varying Pendant Ligands. Inorganic Chemistry, 47(10), 4310-4318. doi:10.1021/ic702393zCervera, J., Schiedt, B., Neumann, R., Mafé, S., & Ramírez, P. (2006). Ionic conduction, rectification, and selectivity in single conical nanopores. The Journal of Chemical Physics, 124(10), 104706. doi:10.1063/1.2179797Ramírez, P., Apel, P. Y., Cervera, J., & Mafé, S. (2008). Pore structure and function of synthetic nanopores with fixed charges: tip shape and rectification properties. Nanotechnology, 19(31), 315707. doi:10.1088/0957-4484/19/31/315707Ali, M., Ramirez, P., Mafé, S., Neumann, R., & Ensinger, W. (2009). A pH-Tunable Nanofluidic Diode with a Broad Range of Rectifying Properties. ACS Nano, 3(3), 603-608. doi:10.1021/nn900039fCervera, J., Alcaraz, A., Schiedt, B., Neumann, R., & Ramírez, P. (2007). Asymmetric Selectivity of Synthetic Conical Nanopores Probed by Reversal Potential Measurements. The Journal of Physical Chemistry C, 111(33), 12265-12273. doi:10.1021/jp071884cVan der Heyden, F. H. J., Bonthuis, D. J., Stein, D., Meyer, C., & Dekker, C. (2007). Power Generation by Pressure-Driven Transport of Ions in Nanofluidic Channels. Nano Letters, 7(4), 1022-1025. doi:10.1021/nl070194hAlcaraz, A., Ramírez, P., García-Giménez, E., López, M. L., Andrio, A., & Aguilella, V. M. (2006). A pH-Tunable Nanofluidic Diode: Electrochemical Rectification in a Reconstituted Single Ion Channel. The Journal of Physical Chemistry B, 110(42), 21205-21209. doi:10.1021/jp063204wAli, M., Nasir, S., Ramirez, P., Cervera, J., Mafe, S., & Ensinger, W. (2012). Calcium Binding and Ionic Conduction in Single Conical Nanopores with Polyacid Chains: Model and Experiments. ACS Nano, 6(10), 9247-9257. doi:10.1021/nn303669