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A study on photolinkers used for biomolecule attachment to polymer surfaces

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Abstract

The use of photolinkers (photoactivatable heterobifunctional crosslinkers) is a popular method to attach biomolecules to polymer surfaces. This study addresses the selection of photolinker and the adjustment of reaction conditions, such as the concentration of biomolecule applied, and irradiation time. The influence of these variables are investigated for four prominent photolinkers: ketyl-reactive benzophenone (BP) and anthraquinone (AQ), nitrene-reactive nitrophenyl azide (NPA), and carbene-reactive phenyl-(trifluoromethyl)diazirine (PTD). The influence of substrate material is discussed, and three different polymers served as representative substrates: poly(methyl methacrylate) (PMMA), polystyrene (PS), and a cycloolefin copolymer (COC). We compared the overall photolinking efficiency of all photolinkers with respect to the polymer substrate they are applied to, and we found considerable differences for certain photolinker/substrate combinations. Of all photolinkers and substrates tested, PTD as photolinker and COC as substrate showed the highest photolinking efficiencies and fastest reaction times. For this study DNA oligonucleotides were chosen as a model system of biomolecular probes, and fluorescence detection of DNA microarrays served as method of detection.

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Abbreviations

AQ:

anthraquinone

BP:

benzophenone

BSA:

bovine serum albumin

COC:

cycloolefin copolymer

E PL :

photolinking efficiency

NPA:

p-nitrophenyl azide

PBS:

phosphate-buffered saline

PL:

photolinker

PMMA:

poly(methyl methacrylate)

PS:

polystyrene

PTD:

3-phenyl-3-(trifluoromethyl)diazirine

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Acknowledgments

The presented work was part of the German national project “nanoMAP: modular application platform with integrated nanoliter dosage for highly parallel sample analysis” supported by the Federal Ministry of Education and Research (project no. 0312001B). The authors are grateful to the project partners microParts (Dortmund, Germany) for providing samples of polymer slides, and IMTEK (Freiburg, Germany) for their support regarding TopSpot technology. Daniela M. Dankbar was financially supported by the DFG Graduiertenkolleg “Analytische Chemie”.

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  1. Daniela M. Dankbar

    Present address: Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO, 80309, USA

Authors and Affiliations

  1. Institute of Physical and Theoretical Chemistry, University of Tübingen, Auf der Morgenstelle 8, 72076, Tübingen, Germany

    Daniela M. Dankbar & Günter Gauglitz

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  1. Daniela M. Dankbar
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  2. Günter Gauglitz
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Correspondence to Günter Gauglitz.

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Dankbar, D.M., Gauglitz, G. A study on photolinkers used for biomolecule attachment to polymer surfaces. Anal Bioanal Chem 386, 1967–1974 (2006). https://doi.org/10.1007/s00216-006-0871-x

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  • DOI: https://doi.org/10.1007/s00216-006-0871-x

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