Controlling the Bioreceptor Spatial Distribution at the Nanoscale for Single Molecule Counting in Microwell Arrays. 2019

Devin Daems, and Iene Rutten, and Jonathan Bath, and Deborah Decrop, and Hans Van Gorp, and Elena Pérez Ruiz, and Steven De Feyter, and Andrew J Turberfield, and Jeroen Lammertyn
Department of Biosystems, Biosensors Group , KU Leuven , Willem de Croylaan 42 , B-3001 Leuven , Belgium.

The ability to detect low concentrations of protein biomarkers is crucial for the early-stage detection of many diseases and therefore indispensable for improving diagnostic devices for healthcare. Here, we demonstrate that by integrating DNA nanotechnologies like DNA origami and aptamers, we can design innovative biosensing concepts for reproducible and sensitive detection of specific targets. DNA origami structures decorated with aptamers were studied as a novel tool to structure the biosensor surface with nanoscale precision in a digital detection bioassay, enabling control of the density, orientation, and accessibility of the bioreceptor to optimize the interaction between target and aptamer. DNA origami was used to control the spatial distribution of an in-house-generated aptamer on superparamagnetic microparticles, resulting in an origami-linked digital aptamer bioassay to detect the main peanut antigen Ara h1 with 2-fold improved signal-to-noise ratio and 15-fold improved limit of detection compared to a digital bioassay without DNA origami. Moreover, the sensitivity achieved was 4 orders of magnitude higher than commercially available and literature-reported enzyme-linked immunosorbent assay techniques. In conclusion, this novel and innovative approach to engineer biosensing interfaces will be of major interest to scientists and clinicians looking for new molecular insights and ultrasensitive detection of a broad range of targets, and, for the next generation of diagnostics.

UI MeSH Term Description Entries
D008958 Models, Molecular Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures. Molecular Models,Model, Molecular,Molecular Model
D009690 Nucleic Acid Conformation The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape. DNA Conformation,RNA Conformation,Conformation, DNA,Conformation, Nucleic Acid,Conformation, RNA,Conformations, DNA,Conformations, Nucleic Acid,Conformations, RNA,DNA Conformations,Nucleic Acid Conformations,RNA Conformations
D000072760 Single Molecule Imaging High resolution imaging techniques that allow visualization of individual molecules of proteins, lipids, or nucleic acids within cells or tissues. Single Molecule Analysis,Single Molecule Tracking,Single Particle Analysis,Single Particle Imaging,Single Particle Microscopy,Single Particle Spectroscopy,Single Particle Tracking,Single Molecule Microscopy,Single Molecule Spectroscopy,Analyses, Single Particle,Analysis, Single Molecule,Analysis, Single Particle,Imaging, Single Molecule,Imaging, Single Particle,Microscopies, Single Particle,Microscopy, Single Molecule,Microscopy, Single Particle,Particle Tracking, Single,Single Molecule Analyses,Single Particle Analyses,Single Particle Microscopies,Single Particle Spectroscopies,Single Particle Trackings,Spectroscopy, Single Molecule,Spectroscopy, Single Particle,Tracking, Single Molecule,Tracking, Single Particle
D001681 Biological Assay A method of measuring the effects of a biologically active substance using an intermediate in vivo or in vitro tissue or cell model under controlled conditions. It includes virulence studies in animal fetuses in utero, mouse convulsion bioassay of insulin, quantitation of tumor-initiator systems in mouse skin, calculation of potentiating effects of a hormonal factor in an isolated strip of contracting stomach muscle, etc. Bioassay,Assay, Biological,Assays, Biological,Biologic Assay,Biologic Assays,Assay, Biologic,Assays, Biologic,Bioassays,Biological Assays
D012825 Silicon A trace element that constitutes about 27.6% of the earth's crust in the form of SILICON DIOXIDE. It does not occur free in nature. Silicon has the atomic symbol Si, atomic number 14, and atomic weight [28.084; 28.086]. Silicon-28,Silicon 28
D052157 Aptamers, Nucleotide Nucleotide sequences, generated by iterative rounds of SELEX APTAMER TECHNIQUE, that bind to a target molecule specifically and with high affinity. DNA Aptamer,DNA Aptamers,RNA Aptamers,Rna Aptamer,Nucleotide Aptamers,Oligonucleotide Ligands, DNA,Oligonucleotide Ligands, RNA,Aptamer, DNA,Aptamer, Rna,Aptamers, DNA,Aptamers, RNA,DNA Oligonucleotide Ligands,RNA Oligonucleotide Ligands
D055616 Microtechnology Manufacturing technology for making microscopic devices in the micrometer range (typically 1-100 micrometers), such as integrated circuits or MEMS. The process usually involves replication and parallel fabrication of hundreds or millions of identical structures using various thin film deposition techniques and carried out in environmentally-controlled clean rooms. Microfabrication,Micromachining,Micromanufacturing,Microfabrications,Micromachinings,Micromanufacturings,Microtechnologies
D036103 Nanotechnology The development and use of techniques to study physical phenomena and construct structures in the nanoscale size range or smaller. Nanotechnologies

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