Fabrication of Oligonucleotide and Protein Arrays on Rigid and Flexible Substrates Coated with Reactive Polymer Multilayers

Broderick, Adam H.; Carter, Matthew C. D.; Lockett, Matthew R.; Smith, Lloyd M.; Lynn, David M.

doi:10.1021/am302285n

Cited by 20 publications

(31 citation statements)

References 69 publications

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“…The signal-to-noise ratios measured here are comparable to those reported for spotted DNA arrays on the same type of substrate. 34 A number of potential applications for RNA arrays, such as the study of RNA binding proteins, aptamers, or the development of new biosensors, depend critically upon minimizing non-specific interactions between the surface and analytes of interest. The ability to synthesize the arrays on a polymeric surface provides an important new avenue for the control of such interactions.…”

Section: Resultsmentioning

confidence: 99%

Photolithographic Synthesis of High-Density DNA and RNA Arrays on Flexible, Transparent, and Easily Subdivided Plastic Substrates

Holden

Carter

et al. 2015

Anal. Chem.

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The photolithographic fabrication of high-density DNA and RNA arrays on flexible and transparent plastic substrates is reported. The substrates are thin sheets of poly(ethylene terephthalate) (PET) coated with crosslinked polymer multilayers that present hydroxyl groups suitable for conventional phosphoramidite-based nucleic acid synthesis. We demonstrate that by modifying MAS procedures to accommodate the physical and chemical properties of these materials, it is possible to synthesize plastic-backed oligonucleotide arrays with feature sizes as small as 14 × 14 μm and feature densities in excess of 125 000/cm2, similar to specifications attainable using rigid substrates such as glass or glassy carbon. These plastic-backed arrays are tolerant to a wide range of hybridization temperatures, and improved synthetic procedures are described that enable the fabrication of arrays with sequences up to 50 nucleotides in length. These arrays hybridize with S/N ratios comparable to those fabricated on otherwise identical arrays prepared on glass or glassy carbon. This platform supports the enzymatic synthesis of RNA arrays and proof-of-concept experiments are presented showing that the arrays can be readily sub-divided into smaller arrays (or ‘millichips’) using common laboratory-scale laser cutting tools. These results expand the utility of oligonucleotide arrays fabricated on plastic substrates and open the door to new potential applications for these important bioanalytical tools.

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Section: Resultsmentioning

confidence: 99%

Photolithographic Synthesis of High-Density DNA and RNA Arrays on Flexible, Transparent, and Easily Subdivided Plastic Substrates

Holden

Carter

et al. 2015

Anal. Chem.

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“…355 Covalent LbL assemblies of PEI and the aminereactive, azlactone-functionalized polymer poly(2-vinyl-4,4-dimethylazlactone) were first prepared on planar glass slides, on which amine-terminated oligonucleotide probe sequences had been manually spotted. Hybridization of fluorescently labeled complementary sequences provided fluorescence spots with high signal intensities, high signal-to-noise ratios, and high sequence specificity.…”

Section: ¹1mentioning

confidence: 99%

Layer-by-layer Nanoarchitectonics: Invention, Innovation, and Evolution

et al. 2013

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Materials fabrication with nanoscale structural precision based on bottom-up-type self-assembly has become more important in various current disciplines in chemistry including materials chemistry, organic chemistry, physical chemistry, analytical chemistry, biochemistry, colloid and surface chemistry, and supramolecular chemistry. Although the design of new materials based on nanoscale self-assembly is anticipated as a key concept, preparing complete three-dimensional structures at nanoscale precision remains a difficult target using current technologies. Rather, dimension-reduced approaches such as layering of two-dimensional nanostructures into precisely controlled lamellar nanomaterials are currently achievable. In particular, layer-by-layer (LbL) assembly is known as a highly versatile method for fabrication of controlled layered structures from various kinds of component materials using very simple, inexpensive, and rapid procedures. Therefore, fabrication of multilayer films through the LbL deposition process has attracted growing interest from various research communities. The high versatility and flexibility of LbL assembly is continuously creating new concepts, new materials, new procedures, and new applications. In this highlight review, we focus on nanoarchitectonics by LbL assembly. After an initial introduction on the invention and a brief history of the LbL assembly technique, innovations and the evolution of the technique are described based mainly on recent examples, which are categorized into two sections: (i) developments in methodology (technical, material, and phenomenological aspects with expansion of concept) and (ii) progress in applications (physical, chemical/biochemical, and biomedical applications).

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“…Past studies from our group have demonstrated that interfacial reactions between amine-reactive poly(2-vinyl-4,4-dimethylazlactone) (PVDMA) and the amine-containing polymer poly(ethylenimine) (PEI) can drive LbL assembly into films that are either thin, smooth, and relatively featureless at the nanometer scale, − or thicker (micrometer-scale) with significant nano- and microscale surface features and substantial nanoscale porosity. − These PEI/PVDMA multilayers are potentially useful in at least three ways: (i) similar to other LbL processes, these materials can be fabricated on a broad range of topologically complex substrates, (ii) the assemblies that result exhibit superior stabilities in complex chemical environments owing to the presence of hydrolytically stable amide-based cross-links, and (iii) the resulting films contain residual azlactone groups that can be used to further functionalize and tailor bulk and surface properties by reactions with a range of nucleophiles . These useful properties make PEI/PVDMA multilayers a versatile platform for the elaboration of a broad range of new functional materials, including tailored biointerfaces, , reactive polymer-based capsules, and new types of chemical and biomolecular sensors . More recently, chemically reactive PEI/PVDMA films with substantial degrees of nanoscale roughness and porosity have made possible the design of surface coatings with extreme wetting behaviors (e.g., superhydrophobicity, underwater superoleophobicity, etc.…”

Section: Introductionmentioning

confidence: 99%

Influence of Side Chain Hydrolysis on the Evolution of Nanoscale Roughness and Porosity in Amine-Reactive Polymer Multilayers

et al. 2020

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We report the influence of side chain hydrolysis on the evolution of nanoscale structure in thin films fabricated by the reactive layer-by-layer (LbL) assembly of branched poly(ethylenimine) (PEI) and poly(2-vinyl-4,4-dimethylazlactone) (PVDMA). LbL assembly of PEI and PVDMA generally leads to the linear growth of thin, smooth films. However, assembly using PVDMA containing controlled degrees of side chain hydrolysis leads to the growth of thicker films that exhibit substantial nanoscale roughness, porosity, and have resulting physicochemical behaviors (e.g., superhydrophobicity) that are similar to those of some thicker PEI/PVDMA coatings reported in past studies. Our results reveal that the degree of PVDMA partial hydrolysis (or carboxylic acid group content) influences the extent to which complex film features develop, suggesting that ion-pairing interactions between hydrolyzed side chains and amines in PEI promote the evolution of bulk and surface morphology. Additional experiments demonstrate that these features likely arise from polymer/polymer interactions at the surfaces of the films during assembly, and not from the formation and deposition of solution-phase polymer aggregates. When combined, our results suggest that nanoporous structures and rough features observed in past studies likely arise, at least in part, from some degree of adventitious side chain hydrolysis in the PVDMA used for film fabrication. Our results provide useful insight into molecular-level features that govern the growth and structures of these reactive materials, and provide a framework to promote nanoscale morphology reliably and reproducibly. The principles and tools reported here should prove useful for further tuning the porosities and tailoring the physicochemical behaviors of these reactive coatings in ways that are important in applied contexts.

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Fabrication of Oligonucleotide and Protein Arrays on Rigid and Flexible Substrates Coated with Reactive Polymer Multilayers

Cited by 20 publications

References 69 publications

Photolithographic Synthesis of High-Density DNA and RNA Arrays on Flexible, Transparent, and Easily Subdivided Plastic Substrates

Photolithographic Synthesis of High-Density DNA and RNA Arrays on Flexible, Transparent, and Easily Subdivided Plastic Substrates

Layer-by-layer Nanoarchitectonics: Invention, Innovation, and Evolution

Influence of Side Chain Hydrolysis on the Evolution of Nanoscale Roughness and Porosity in Amine-Reactive Polymer Multilayers

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