In situ fabrication of cleavable peptide arrays on polydimethylsiloxane and applications for kinase activity assays

Chen, Huang Han; Hsiao, Yu Chieh; Li, Jie Ren; Chen, Shu Hui

doi:10.1016/j.aca.2015.01.040

Cited by 6 publications

(4 citation statements)

References 21 publications

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“…6,7 Compared with inorganic support, polymer substrates show good processability and are lightweight and impact resistant; thus, they are conducive to large-scale industrial production of low-cost disposable biochips. Recently, many polymer substrate such as poly(methyl methacrylate) (PMMA), 8,9 polystyrene (PS), 10,11 polycarbonate (PC), 12,13 poly(ethylene terephthalate) (PET), 14 polydimethylsiloxane (PDMS), 15,16 and cyclic olefin copolymer (COC) 17,18 have been investigated as substrates for biochip preparation.…”

Section: Introductionmentioning

confidence: 99%

“…Protein microarrays have become a powerful tool in biorelated fields such as precision diagnostics, biomarker screening, drug screening, and proteomics due to the advantages of high throughput, miniaturization, fast quantification, convenient handling, and reduced amount of analyte. − Inorganic materials such as glass and silicon wafer have been widely used as a substrate for the fabrication of biochips because of their excellent bulk properties and surface reactiveness for further modification. , Compared with inorganic support, polymer substrates show good processability and are lightweight and impact resistant; thus, they are conducive to large-scale industrial production of low-cost disposable biochips. Recently, many polymer substrate such as poly(methyl methacrylate) (PMMA), , polystyrene (PS), , polycarbonate (PC), , poly(ethylene terephthalate) (PET), polydimethylsiloxane (PDMS), , and cyclic olefin copolymer (COC) , have been investigated as substrates for biochip preparation.…”

Section: Introductionmentioning

confidence: 99%

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Facile Surface Functionalization of Cyclic Olefin Copolymer Film with Anhydride Groups for Protein Microarray Fabrication

Yuan

Wang

Chen

et al. 2020

ACS Appl. Bio Mater.

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Immobilization of protein at high efficiency is a challenge for fabricating polymer-based protein chips. Here, a simple but effective approach was developed to fabricate a cyclic olefin copolymer (COC)-based protein microarray with a high immobilization density. In this strategy, poly(maleic anhydride-co-vinyl acetate) (poly(MAH-co-VAc)) brushes were facilely attached on the COC surface via UV-induced graft copolymerization. The introduction of poly(MAH-co-VAc) brushes resulted in an obvious increase in the surface roughness of COC. The functionalized COC showed little reduction in transparency compared with pristine COC, indicating that the photografting treatment did not alter its optical property. The graft density of the anhydride groups on the modified COC could be tuned from 0.46 to 3.2 μmol/cm2. The immobilization efficiency of immunoglobulin G (IgG) on functionalized COC reached 88% due to the high reactivity between anhydride groups and amine groups of IgGs. An immunoassay experiment demonstrated that the microarray showed high sensitivity to the target analyte.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Facile Surface Functionalization of Cyclic Olefin Copolymer Film with Anhydride Groups for Protein Microarray Fabrication

Yuan

Wang

Chen

et al. 2020

ACS Appl. Bio Mater.

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“…The area of contact between the substrate and the PDMS stamp also serves as a structural template to guide the formation of NP assemblies with designed geometry. During the pattern transfer process, free residual Si–H groups of the PMDS stamp surface serve as the reducing agent to gradually reduce Au or Ag ions, forming Au or Ag NPs at ambient conditions (Figure C). − After the process is completed, the PDMS stamp is carefully peeled off. A single layer of close-packed Au or Ag particles mixed with few PDMS residues is synthesized and transferred from the protruding surface of the designed PDMS stamp, simultaneously (Figure D).…”

Section: Resultsmentioning

confidence: 99%

Hierarchical Nanoparticle Assemblies Formed via One-Step Catalytic Stamp Pattern Transfer

Hung

Liu

Lee

et al. 2019

ACS Appl. Mater. Interfaces

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The one-step catalytic stamp pattern transfer process is described for producing arrays of hierarchical nanoparticle assemblies. The method simply combines in situ nanoparticle synthesis triggered by free residual Si−H groups on PDMS stamps and the lift-off pattern transfer technique. No additional nanoparticle synthesis procedure is required before the pattern transfer process. Exquisitely uniform and precisely spaced hierarchical nanoparticle assemblies with designed geometry can be rapidly produced using the catalytic stamp pattern transfer process. Sequential catalytic stamp pattern transfer also is described to generate multilayered, hierarchical nanoparticle assemblies with various geometries. The hierarchical nanoparticle assemblies catalytically transferred onto the surface are not just nanoparticles but nanoparticle− polydimethylsiloxane residue composites. The in situ-synthesized nanoparticles retain optical properties. The hierarchical nanoparticle assemblies with precisely controlled geometry further show potential in the application of surface-enhanced Raman scattering. The capability of one-step catalytic stamp pattern transfer allows the scalable and reproducible fabrication of welldefined hierarchical nanoparticle assemblies.

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“…24 But the most common are disulfide linkers, which are easily cleaved by reductive agents. They were used to immobilize peptides and proteins on planar substrates, 25,26 nanoparticles, 27 hydrogels, 28 or for release of fluorophores from the surface into solution in a cleavable tag immunoassay. 29 However, to our knowledge, cleavable linkers have never been used in microarray blocking agents before.…”

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confidence: 99%

Improving Immunoassay Performance with Cleavable Blocking of Microarrays

2020

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Among the key issues that are commonly associated with the development of microarray-based assays are nonspecific binding and diffusion constraints. Here we present a novel strategy addressing both of these challenges simultaneously. The essence of the method consists in blocking the microarray surface with a blocking agent containing a perfluoroalkyl chain and a disulfide linker. The resulting surface is hydrophobic, and no immiscible liquid layer remains on it upon cyclically draining and replenishing the sample solution, ensuring an efficient mass transfer of an analyte onto a microarray. Prior to the signal detection procedure, disulfide bonds are chemically cleaved, and the perfluoroalkyl chains are removed from the microarray surface along with nonspecifically adsorbed proteins, resulting in extremely low background. Using conventional fluorescent detection, we show a 30-fold increase in signal/background ratio compared to a common epoxy-modified glass substrate. The combination of this technique with magnetic beads detection results in a simple and ultrasensitive cholera toxin (CT) immunoassay. The limit of detection (LOD) is 1 fM, which is achieved with an analyte binding time of 1 h. Efficient mass transfer provides highly sensitive detection of whole virus particles despite their low diffusion coefficient. The achieved LOD for vaccinia virus is 10 4 particles in 1 mL of sample. Finally, we have performed for the first time the simultaneous detection of whole virus and CT protein biomarker in a single assay. The developed technique can be used for multiplex detection of trace amounts of pathogens of various natures.

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In situ fabrication of cleavable peptide arrays on polydimethylsiloxane and applications for kinase activity assays

Cited by 6 publications

References 21 publications

Facile Surface Functionalization of Cyclic Olefin Copolymer Film with Anhydride Groups for Protein Microarray Fabrication

Facile Surface Functionalization of Cyclic Olefin Copolymer Film with Anhydride Groups for Protein Microarray Fabrication

Hierarchical Nanoparticle Assemblies Formed via One-Step Catalytic Stamp Pattern Transfer

Improving Immunoassay Performance with Cleavable Blocking of Microarrays

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