Facile Photoimmobilization of Proteins onto Low-Binding PEG-Coated Polymer Surfaces

Esben Kjær Unmack Larsen, Morten Bo Lindholm Mikkelsen, Niels Bent Larsen

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Immobilization of proteins onto polymer surfaces usually requires specific reactive functional groups. Here, we show an easy one-step method to conjugate protein covalently onto almost any polymer surface, including low protein-binding poly(ethylene glycol) (PEG), without the requirement for the presence of specific functional groups. Several types of proteins, including alkaline phosphatase, bovine serum albumin, and polyclonal antibodies, were photoimmobilized onto a PEG-coated polymer surface using a water-soluble benzophenone as photosensitizer. Protein functionality after immobilization was verified for both enzymes and antibodies, and their presence on the surface was confirmed by X-ray photoelectron spectroscopy (XPS) and confocal fluorescence microscopy. Conjugation of capture antibody onto the PEG coating was employed for a simplified ELISA protocol without the need for blocking uncoated surface areas, showing ng/mL sensitivity to a cytokine antigen target. Moreover, spatially patterned attachment of fluorescently labeled protein onto the low-binding PEG-coated surface was achieved with a projection lithography system that enabled the creation of micrometer-sized protein features.
    Original languageEnglish
    JournalBiomacromolecules
    Volume15
    Issue number3
    Pages (from-to)894-899
    ISSN1526-4602
    DOIs
    Publication statusPublished - 2014

    Cite this

    Larsen, Esben Kjær Unmack ; Mikkelsen, Morten Bo Lindholm ; Larsen, Niels Bent. / Facile Photoimmobilization of Proteins onto Low-Binding PEG-Coated Polymer Surfaces. In: Biomacromolecules. 2014 ; Vol. 15, No. 3. pp. 894-899.
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    title = "Facile Photoimmobilization of Proteins onto Low-Binding PEG-Coated Polymer Surfaces",
    abstract = "Immobilization of proteins onto polymer surfaces usually requires specific reactive functional groups. Here, we show an easy one-step method to conjugate protein covalently onto almost any polymer surface, including low protein-binding poly(ethylene glycol) (PEG), without the requirement for the presence of specific functional groups. Several types of proteins, including alkaline phosphatase, bovine serum albumin, and polyclonal antibodies, were photoimmobilized onto a PEG-coated polymer surface using a water-soluble benzophenone as photosensitizer. Protein functionality after immobilization was verified for both enzymes and antibodies, and their presence on the surface was confirmed by X-ray photoelectron spectroscopy (XPS) and confocal fluorescence microscopy. Conjugation of capture antibody onto the PEG coating was employed for a simplified ELISA protocol without the need for blocking uncoated surface areas, showing ng/mL sensitivity to a cytokine antigen target. Moreover, spatially patterned attachment of fluorescently labeled protein onto the low-binding PEG-coated surface was achieved with a projection lithography system that enabled the creation of micrometer-sized protein features.",
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    Facile Photoimmobilization of Proteins onto Low-Binding PEG-Coated Polymer Surfaces. / Larsen, Esben Kjær Unmack; Mikkelsen, Morten Bo Lindholm; Larsen, Niels Bent.

    In: Biomacromolecules, Vol. 15, No. 3, 2014, p. 894-899.

    Research output: Contribution to journalJournal articleResearchpeer-review

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    AU - Larsen, Esben Kjær Unmack

    AU - Mikkelsen, Morten Bo Lindholm

    AU - Larsen, Niels Bent

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    AB - Immobilization of proteins onto polymer surfaces usually requires specific reactive functional groups. Here, we show an easy one-step method to conjugate protein covalently onto almost any polymer surface, including low protein-binding poly(ethylene glycol) (PEG), without the requirement for the presence of specific functional groups. Several types of proteins, including alkaline phosphatase, bovine serum albumin, and polyclonal antibodies, were photoimmobilized onto a PEG-coated polymer surface using a water-soluble benzophenone as photosensitizer. Protein functionality after immobilization was verified for both enzymes and antibodies, and their presence on the surface was confirmed by X-ray photoelectron spectroscopy (XPS) and confocal fluorescence microscopy. Conjugation of capture antibody onto the PEG coating was employed for a simplified ELISA protocol without the need for blocking uncoated surface areas, showing ng/mL sensitivity to a cytokine antigen target. Moreover, spatially patterned attachment of fluorescently labeled protein onto the low-binding PEG-coated surface was achieved with a projection lithography system that enabled the creation of micrometer-sized protein features.

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