Site-Selective, Cleavable Linkers: Quality Control and the Characterization of Small Molecules on Microelectrode Arrays

Bi, Bo; Huang, Richard Y.‐C.; Maurer, Karl; Chen, Ceng; Moeller, Kevin D.

doi:10.1021/jo2017907

Cited by 14 publications

(16 citation statements)

References 34 publications

(31 reference statements)

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“…While this collection of challenges initially seems daunting, modern techniques in electroorganic synthesis offer a powerful means for addressing each of them. Mediated electrochemical reactions can be used to place molecules by specific electrodes in the array, modify the molecules once they are on the array, cleave the molecules from the surface for characterization, modify the surface of the array in ways that allow for optimization of both the synthesis and analytical studies needed, and then control the nature of the resulting analytical surface using reversible reactions. The result is a potentially powerful new platform for screening molecular libraries that retains an unprecedented level of synthetic control, and hence quality control, over the molecular library.…”

Section: Discussionmentioning

confidence: 99%

Electroorganic Synthesis and the Construction of Addressable Molecular Surfaces

2019

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In this Minireview, an electroorganic synthesis approach to the construction of addressable, complex molecular surfaces is described along with the parameters that guided the development of that synthetic approach. The result of the work is a synthetic toolbox that will allow microelectrode arrays to be used for the “real‐time” monitoring of small molecule interactions with biological targets.

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Section: Discussionmentioning

confidence: 99%

Electroorganic Synthesis and the Construction of Addressable Molecular Surfaces

2019

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“…A subsequent cyclization reaction to form a lactone cleaves the molecule from the surface. For the arrays, the safety-catch linker used has taken advantage of a t- Boc protected alcohol (XP = O t- Boc) that could be cleaved by generating acid at a selected electrode …”

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Microelectrode Arrays, Dihydroxylation, and the Development of an Orthogonal Safety-Catch Linker

2021

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Construction of larger molecular libraries on an addressable microelectrode array requires a method for recovering and characterizing molecules from the surface of any electrode in the array. This method must be orthogonal to the synthetic strategies needed to build the array. We report here a method for achieving this goal that employs the site-selective dihydroxylation reaction of a simple olefin.

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“…Photochemical cleavage of the PEG coating on a self-assembled monolayer was able to control cell adhesion . Covalent immobilization of DNA or proteins via cleavable linkers allowed immobilized molecules to be released from the surface by UV irradiation, under the action of the enzyme, base, or acid . But the most common are disulfide linkers, which are easily cleaved by reductive agents.…”

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confidence: 99%

“…20 Covalent immobilization of DNA or proteins via cleavable linkers allowed immobilized molecules to be released from the surface by UV irradiation, 21 under the action of the enzyme, 22 base, 23 or acid. 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.…”

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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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Site-Selective, Cleavable Linkers: Quality Control and the Characterization of Small Molecules on Microelectrode Arrays

Cited by 14 publications

References 34 publications

Electroorganic Synthesis and the Construction of Addressable Molecular Surfaces

Electroorganic Synthesis and the Construction of Addressable Molecular Surfaces

Microelectrode Arrays, Dihydroxylation, and the Development of an Orthogonal Safety-Catch Linker

Improving Immunoassay Performance with Cleavable Blocking of Microarrays

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