Abstract
Defined arrangements of individual molecules are covalenty connected ("printed") onto SAM-functionalised gold substrates with nanometer resolution. Substrates were initially pre-functionlised by coating with 3,3'-dithiodipropionic acid (DTPA) to form a self-assembled monolayer (SAM), which was characterised by atomic force microscopy (AFM), contact angle goniometry, cyclic voltammetry and surface plasmon resonance (SPR) spectroscopy. Pre-defined "ink" patterns displayed on DNA origami-based single-use carriers ("stamp") were covalently conjugated to the SAM using 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) and N-hydroxy-succinimide (NHS). These anchor points were used to create nanometer-precise single-molecule arrays, here with complementary DNA and streptavidin. Sequential steps of the printing process were evaluated by AFM and SPR spectroscopy. It was shown that 30% of the detected arrangements closely match the expected length distribution of designed patterns, whereas another 40% exhibit error within the range of only 1 streptavidin molecule. SPR results indicate that imposing a defined separation between molecular anchor points within the pattern through this printing process enhances the efficiency for association of specific binding partners for systems with high sterical hindrance. This study expands upon earlier findings where geometrical information was conserved by the application of DNA nanostructures, by establishing a generalisable strategy which is universally applicable to nearly any type of prefunctionalised substrate such as metals, plastics, silicates, ITO or 2D materials.
Highlights
Current trends in miniaturisation for the fabrication of microcircuitry, sensing devices and array-based analytical systems amplify the demand for precise and cost-effective enabling technologies for nanoscale surface lithography.[1]
In order to universalise the protocol for printing patterns of ink molecules positioned on DNA origami stamps, targeted substrates first need to be pre-functionalised
Bioconjugation of DNA strands to the self-assembled monolayer (SAM)-coated surface was facilitated by forming amide-bonds between the carboxylic acid groups on the dithiodipropionic acid (DTPA) molecules and amine groups on the ink strands
Summary
Current trends in miniaturisation for the fabrication of microcircuitry, sensing devices and array-based analytical systems amplify the demand for precise and cost-effective enabling technologies for nanoscale surface lithography.[1]. M13 bacteriophage genome and around 7000 bases long, which allows it to be folded by the approximately 200 staple strands into nearly any 2D or 3D structure, with typical dimensions on the order of tens or hundreds of nanometers This technique offers the unmatched possibility to deterministically program the shape and surface functionality of nanostructures that selfassemble in a scalable, high-yield assembly reaction, and the possibility to achieve sub-nanometre spatial resolution for individual molecules.[6] This elegant method enables the positioning and detailed study of individual nano-objects e.g. biomolecules,[7] nanoparticles[8] or dyes[9] in a straightforward manner with high precision via a simple breadboard system. A carboxylic acid group facilitated the printing of common amine-labeled DNA strands, arranged in distinct patterns by a DNA origami carrier
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