Nanoscale Protein Patterning by Imprint Lithography

Hoff, J. Damon; Cheng, Li-Jing; Meyhöfer, Edgar; Guo, L. Jay; Hunt, Alan

doi:10.1021/nl049758x

Cited by 288 publications

(223 citation statements)

References 20 publications

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“…Our method can be modified slightly to accommodate these fabrication methods by derivatizing the glass surface after the fabrication of the wells to prevent the destruction of the biotin moieties during the imprinting or etching steps. [32][33][34] With the densities we have shown here, more than 20 million beads can be arrayed in 1 cm 2 . We have also demonstrated the ability to fabricate arrays of wells with dimensions as small as 0.8 μm and densities approaching 40 million wells/cm 2 over a large area on a cover glass with the stepper system we used.…”

Section: Resultsmentioning

confidence: 68%

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Magnetic Assembly of High-Density DNA Arrays for Genomic Analyses

Barbee

Huang

2008

Anal. Chem.

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A method for rapidly assembling high-density DNA arrays with near-perfect order is described. Photolithography is used to generate a wafer-scale array of microwells in a layer of photoresist on a chemically functionalized glass coverslip. The array is enclosed within a microfluidic device, and a suspension of superparamagnetic microbeads conjugated to DNA molecules is introduced into the chamber. A permanent magnet is used to direct the rapid assembly of the beads into the wells, with each well containing a single bead. These beads are immobilized on the glass surface via affinity binding, and excess beads can be recycled or washed away. Nonspecifically bound beads are removed by dissolving the photoresist. The result is a high-density array of beads with virtually no background. This method can be used to produce protein arrays for chip-based assays and DNA arrays for genotyping or genome sequencing.Some of the greatest breakthroughs in biomedical research can be attributed to the development of the numerous high-throughput technologies for quantitative measurements of biomolecules. Many of these technologies are made possible by microfabrication techniques commonly used in the semiconductor industry. For example, DNA and protein arrays fabricated by robotic printing and photolithographic methods have enabled extremely large-scale surveys of biomolecules. [1][2][3][4] The emerging "next generation" genome sequencing technologies, many of which utilize massive parallelization and miniaturization to achieve unprecedented multiplexing, throughput and cost reductions, [5][6][7][8][9][10] promise to revolutionize biomedical research and enable personalized healthcare. However, some of these technology platforms utilize randomly distributed DNA-conjugated microbeads or clones on a glass slide within a reaction chamber. The random arrangements of the beads or clones result in low throughput and imaging efficiency, complicated image processing, and high reagent costs. 6-10 One approach to dramatically improve these devices involves the use of microfabricated arrays to eliminate overlap and to minimize the area between the beads or clones.Such arrays can be generated by depositing samples onto glass slides using robotic contact printing, 2 microcontact printing, 11-13 or dip pen lithography. 14 These arrays can also be generated by assembling beads onto microfabricated arrays of wells on glass or silicon substrates [15][16][17][18] or in etched wells on the face of a fiber-optic bundle. 19,20 Since bead assembly will not occur in an efficient and reliable manner if the process depends solely upon gravitational forces and Brownian motion, this process is typically achieved via solvent evaporation or dewetting. [17][18][19][20][21] However, these approaches are not suitable when rapid assembly is required or sample drying is undesirable. Other groups have employed electric 22 and magnetic [23][24][25] assembly methods to overcome these issues, but these active approaches require multistep fabrication processes and...

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Section: Resultsmentioning

confidence: 68%

Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

Magnetic Assembly of High-Density DNA Arrays for Genomic Analyses

Barbee

Huang

2008

Anal. Chem.

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“…Using predominantly non-contact printers based on ink-jet technology, antibody microarrays have been produced and applied having an overall foot print of <1 cm 2 , based on 18 × 10 3 μm 2 (Ø ∼ 150 μm)-sized spots at a density of ⩽2000 spots cm −2 ; and a multiplicity of <850 different antibodies/array [5][6][7]13]. In an effort to evolve the array platform even further with respect to both spot density (>100 000 spots cm −2 ) and spot size (<0.8 μm 2 , Ø < 1 μm), the first conceptual antibody (protein) nanoarrays have been produced [14][15][16][17][18][19][20][21][22][23]; for review see [24]. A set of technologies are available for producing such nanoarrays, e.g.…”

Section: Introductionmentioning

confidence: 99%

“…A set of technologies are available for producing such nanoarrays, e.g. atomic force microscopy (AFM) [25], dip-pen nanolithography (DPN) [26], nanofountain probe [27,28], nanoimprint lithography [16], and various nanodispensers [20,22,29]. Albeit successful, these nanoarray layouts have been found to be associated with a set of key methodological shortcomings.…”

Section: Introductionmentioning

confidence: 99%

Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer

et al. 2014

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Miniaturized (Ø 10 μm), multiplexed (>5-plex), and high-density (>100 000 spots cm−2) antibody arrays will play a key role in generating protein expression profiles in health and disease. However, producing such antibody arrays is challenging, and it is the type and range of available spotters which set the stage. This pilot study explored the use of a novel microspotting tool, BioplumeTM—consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels—to produce miniaturized, multiplexed, and high-density planar recombinant antibody arrays for protein expression profiling which targets crude, directly labelled serum. The results demonstrated that 16-plex recombinant antibody arrays could be produced—based on miniaturized spot features (78.5 um2, Ø 10 μm) at a 7–125-times increased spot density (250 000 spots cm−2), interfaced with a fluorescent-based read-out. This prototype platform was found to display adequate reproducibility (spot-to-spot) and an assay sensitivity in the pM range. The feasibility of the array platform for serum protein profiling was outlined.

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“…monolayers (SAMs) 9,10 or adsorbed copolymers 8 which permit patterning of proteins in the sub-100 nm size range. Another imprint method that has shown utility in the patterning of functional materials is nanocontact molding (NCM).…”

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confidence: 99%

Chain-End Functionalized Nanopatterned Polymer Brushes Grown via in Situ Nitroxide Free Radical Exchange

et al. 2008

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The patterning of biologically active materials has been accomplished by the use of imprint lithography of functional photopolymer resins to create controlled nanoscale patterns of a cross-linked photopolymer containing embedded initiator groups. Functionalized polymer brushes consisting of polystyrene and poly(N,N-dimethylacrylamide) were grown from these patterned layers by nitroxide-mediated polymerization. Chain-end functionalization of the brush layer was accomplished by nitroxide radical exchange during the polymerization. Accordingly, brush layers terminated by pyrene and biotin functional groups were obtained by exchange with the appropriate alkoxyamines. The presence of pyrene functionality at the chain ends of the brushes was confirmed by fluorescent emission measurements. Fluorescently labeled streptavidin protein was selectively attached with high selectivity to the patterned biotinylated brush layer through biotin−streptavidin interactions. The functionalized polymer grafted surfaces and nanopatterns have been successfully characterized using a fluorescence spectrophotometer, AFM, SEM, confocal microscopy, and water contact angle measurements.

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Nanoscale Protein Patterning by Imprint Lithography

Cited by 288 publications

References 20 publications

Magnetic Assembly of High-Density DNA Arrays for Genomic Analyses

Magnetic Assembly of High-Density DNA Arrays for Genomic Analyses

Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer

Chain-End Functionalized Nanopatterned Polymer Brushes Grown via in Situ Nitroxide Free Radical Exchange

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