| Publikationstyp: | Beitrag in wissenschaftlicher Zeitschrift |
| Art der Begutachtung: | Peer review (Publikation) |
| Titel: | A microwell array platform for picoliter membrane protein assays |
| Autor/-in: | Binkert, Andreas Studer, Philipp Vörös, Janos |
| DOI: | 10.1002/smll.200801289 |
| Erschienen in: | Small |
| Band(Heft): | 5 |
| Heft: | 9 |
| Seite(n): | 1070 |
| Seiten bis: | 1077 |
| Erscheinungsdatum: | 2009 |
| Verlag / Hrsg. Institution: | Wiley |
| ISSN: | 1613-6810 1613-6829 |
| Sprache: | Englisch |
| Schlagwörter: | Biological Assay; Biomimetic Material; Lipid Bilayer; Membrane Protein; Microchemistry; Nanostructure; Nanotechnology; Protein Array Analysis |
| Fachgebiet (DDC): | 540: Chemie 615: Pharmakologie und Therapeutik |
| Zusammenfassung: | A novel microwell chip is developed that can be used to detect protein binding in a liquid environment, together with a liquid handling system that allows the performance of assays with picoliter volumes. A PDMS well structure is cast on a planar optical waveguide, providing reaction containers combined with a high-sensitivity fluorescence readout system. Individual wells of the array can be addressed, filled, and rinsed using a contact-mode pin and ring spotter. This allows for immunoassays in a heavily multiplexed way, as all steps of the assay can be individually chosen per well. An array density of over 1000 wells cm(-2) is used for the current experiments. The wells provide a protected liquid environment in which the handling of proteins in their natural state is possible, thus maintaining their activity. The membrane protein annexin V is chosen as a model protein to demonstrate the current possibilities. Annexin V binds to phosphatidylserine (PS) head groups of lipids in a Ca(2+)-dependent manner and is often chosen as a marker for cell apoptosis. Lipid vesicles with and without PS are spotted in individual wells and spontaneously formed a planar lipid bilayer on the bottom of the buffer-filled wells. Annexin V can be used to distinguish between wells containing PS groups previously incorporated in the membrane patches and reference wells without PS head groups. Also, the dependence on the calcium concentration can be shown. Fluorescence readout of the assays is performed using a highly sensitive system based on a planar optical waveguide. |
| URI: | https://digitalcollection.zhaw.ch/handle/11475/1733 |
| Volltext Version: | Publizierte Version |
| Lizenz (gemäss Verlagsvertrag): | Lizenz gemäss Verlagsvertrag |
| Departement: | School of Engineering |
| Enthalten in den Sammlungen: | Publikationen School of Engineering |
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Binkert, A., Studer, P., & Vörös, J. (2009). A microwell array platform for picoliter membrane protein assays. Small, 5(9), 1070–1077. https://doi.org/10.1002/smll.200801289
Binkert, A., Studer, P. and Vörös, J. (2009) ‘A microwell array platform for picoliter membrane protein assays’, Small, 5(9), pp. 1070–1077. Available at: https://doi.org/10.1002/smll.200801289.
A. Binkert, P. Studer, and J. Vörös, “A microwell array platform for picoliter membrane protein assays,” Small, vol. 5, no. 9, pp. 1070–1077, 2009, doi: 10.1002/smll.200801289.
BINKERT, Andreas, Philipp STUDER und Janos VÖRÖS, 2009. A microwell array platform for picoliter membrane protein assays. Small. 2009. Bd. 5, Nr. 9, S. 1070–1077. DOI 10.1002/smll.200801289
Binkert, Andreas, Philipp Studer, and Janos Vörös. 2009. “A Microwell Array Platform for Picoliter Membrane Protein Assays.” Small 5 (9): 1070–77. https://doi.org/10.1002/smll.200801289.
Binkert, Andreas, et al. “A Microwell Array Platform for Picoliter Membrane Protein Assays.” Small, vol. 5, no. 9, 2009, pp. 1070–77, https://doi.org/10.1002/smll.200801289.
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