Journal article
Facile Photoimmobilization of Proteins onto Low-Binding PEG-Coated Polymer Surfaces
Department of Micro- and Nanotechnology, Technical University of Denmark1
Amphiphilic Polymers in Biological Sensing, Department of Micro- and Nanotechnology, Technical University of Denmark2
Polymer Microsystems for Cell Processing, Department of Micro- and Nanotechnology, Technical University of Denmark3
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.
Language: | English |
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Year: | 2014 |
Pages: | 894-899 |
ISSN: | 15264602 and 15257797 |
Types: | Journal article |
DOI: | 10.1021/bm401745a |
ORCIDs: | Larsen, Niels Bent |