Electrokinetic characterization of hybrid NOA 81-glass microchips: Application to protein microchip electrophoresis with indirect fluorescence detection |
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Authors: | Menel Ben Frej Fanny d'Orlyé Gerson F Duarte-Junior Wendell K T Coltro Anne Varenne |
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Institution: | 1. Institute of Chemistry for Life and Health Sciences i-CLeHS, Chimie ParisTech-PSL/CNRS 8060
, Paris, France;2. Instituto de Química, Universidade Federal de Goiás, Goiânia, Brazil |
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Abstract: | A low-cost and straightforward hybrid NOA (Norland optical adhesive) 81-glass microchip electrophoresis device was designed and developed for protein separation using indirect fluorescence detection. This new microchip was first characterized in terms of surface charge density via electroosmotic mobility measurement and stability over time. A systematic determination of the electroosmotic mobility (μeo) over a wide pH range (2–10) and at various ionic strengths (20–50 mM) was developed for the first time via the neutral marker approach in an original simple frontal methodology. The evolution of μeo was proved consistent with the silanol and thiol functions arising from the glass and the NOA materials, respectively. The repeatability and reproducibility of the measurements on different microchips (RSD < 14%) and within 15 days (less than 5% decrease) were successfully demonstrated. The microchip was then applied for the efficient electrophoretic separation of proteins in a zonal mode coupled with indirect fluorescence detection, which is, to our knowledge, the first proof of concept of capillary zone electrophoresis in this hybrid microsystem. |
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Keywords: | electroosmotic mobility hybrid glass-based chip indirect fluorescence detection Norland 81 proteins |
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