On-chip immunoprecipitation for protein purification

Sandison, Mairi E.; Cumming, Sarah A.; Kölch, Walter; Pitt, Andrew R.

doi:10.1039/c005295g

Cited by 23 publications

(22 citation statements)

References 39 publications

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“…The mold used for casting this channel was fabricated as previously described. 29 The scale bar in photograph (d) is 3 mm. Micrograph (e) shows a magnified region of the serpentine channel packed with 50 mm diameter internal pillars, the scale bar is 100 mm.…”

Section: Enrichment Of Phosphopeptidesmentioning

confidence: 99%

“…20,21 For phosphopeptide analysis by MALDI-MS, several reports have described on-target approaches to enrichment. [22][23][24][25] The implementation of phosphopeptide enrichment in a lab-on-a-chip (LOC) format 26 has many potential advantages including low sample volume requirements, the potential both to multiplex parallel analysis streams and integrate several sample preparation stages in a single system, [27][28][29][30][31] decreased analysis times, increased experimental throughput and minimal sample handling.…”

Section: Introductionmentioning

confidence: 99%

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Magnetite-doped polydimethylsiloxane (PDMS) for phosphopeptide enrichment

Sandison

Jensen

Gesellchen

et al. 2014

Analyst

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Reversible phosphorylation plays a key role in numerous biological processes. Mass spectrometry-based approaches are commonly used to analyze protein phosphorylation, but such analysis is challenging, largely due to the low phosphorylation stoichiometry. Hence, a number of phosphopeptide enrichment strategies have been developed, including metal oxide affinity chromatography (MOAC). Here, we describe a new material for performing MOAC that employs a magnetite-doped polydimethylsiloxane (PDMS), that is suitable for the creation of microwell array and microfluidic systems to enable low volume, high throughput analysis. Incubation time and sample loading were explored and optimized and demonstrate that the embedded magnetite is able to enrich phosphopeptides. This substrate-based approach is rapid, straightforward and suitable for simultaneously performing multiple, low volume enrichments.

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Section: Enrichment Of Phosphopeptidesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Magnetite-doped polydimethylsiloxane (PDMS) for phosphopeptide enrichment

Sandison

Jensen

Gesellchen

et al. 2014

Analyst

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“…Even valid, this approach is reversible, and therefore PDMS recovers its hydrophobicity with time. An alternative is to introduce hydroxyl (-OH) groups on the PDMS surface by physisorption of polyvinyl alcohol (PVA) polymer [16] and further silanization [17] of the resulting surface. At this stage, functional groups can be introduced enabling the covalent attachment of biomolecules [18], finally providing to the LoC the desired selectivity for specific analyte while minimizing interference due to non-specific adsorption of other analytes.…”

Section: Introductionmentioning

confidence: 99%

Modular Optofluidic Systems (MOPS)

Ackermann

Dietvorst

Sanchís³

et al. 2016

SPIE Proceedings

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Elementary PDMS-based building blocks of fluidic, optical and optofluidic components for Lab on a chip (LOC) platforms has here been developed. All individual modules are compatible and can be anchored and released with the help of puzzle-type connectors This approach is a powerful toolbox to create modular optofluidic systems (MOPS), which can be modified/upgraded to user needs and in-situ reconfigurable. In addition, the PDMS can locally be functionalized, defining a modular biosensor. Measurements in absorbance and fluorescence have been pursued as demonstrator.

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“…They immobilized glucose oxidase on a PDMS layer after a hydrophilization stepusing a plasma process and further silanization, with the aim of fabricating a glucose sensor. Sandison et al made also use of a plasma andsilanization process to immobilize antibodies on a PDMS column for protein purification applications 12 . But this process also has somedrawbacks: the modification is temporal because the plasma oxidized surface progressively recovers its hydrophobicity.…”

Section: Introductionmentioning

confidence: 99%

Biofunctionalization of PDMS-based microfluidic systems

Ibarlucea¹,

Fernández‐Sánchez²,

Demming

et al. 2011

Protocol Exchange

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Three simple approaches for the selective immobilization of biomolecules on the surface of poly(dimethylsiloxane) (PDMS) microfluidic systems that do not require any specific instrumentation, are described and compared. They are based in the introduction of hydroxyl groups on the PDMS surface by direct adsorption of either polyethylene glycol (PEG) or polyvinyl alcohol (PVA) as well as by a liquid-based oxidation step. The hydroxyl groups are then silanized using a silane containing an aldehyde end-group that allows the surface to interact with a primary amine moiety of the biomolecule structure to be immobilized. The entire process takes 4.5h. The required steps can be characterized in less than 15 hours by contact angle measurements, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The performance of the biofunctionalization process can be assessed by using peroxidase enzyme as a model biomolecule. Its correct immobilization and stability is easily tested by developing an analytical approach for hydrogen peroxide (H2O2) detection in the biofunctionalized microfluidic system and carrying out analytical measurements for a period of up to two months.

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On-chip immunoprecipitation for protein purification

Cited by 23 publications

References 39 publications

Magnetite-doped polydimethylsiloxane (PDMS) for phosphopeptide enrichment

Magnetite-doped polydimethylsiloxane (PDMS) for phosphopeptide enrichment

Modular Optofluidic Systems (MOPS)

Biofunctionalization of PDMS-based microfluidic systems

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