Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies
| Autor: | Jaho, Sofia, Junius, Niels, Borel, Franck, Sallaz-Damaz, Yoann, Salmon, Jean-Baptiste, Budayova-Spano, Monika |
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| Přispěvatelé: | Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Laboratoire du Futur (LOF), Université Sciences et Technologies - Bordeaux 1 (UB)-RHODIA-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), french-CRG beamline BM30A-FIP at ESRF, European Project: RAMP, Université Sciences et Technologies - Bordeaux 1-RHODIA-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | Journal of visualized experiments : JoVE Journal of visualized experiments : JoVE, 2021, 170, ⟨10.3791/61660⟩ 'Journal of Visualized Experiments ', vol: 170, pages: e61660-1-e61660-24 (2021) Journal of visualized experiments : JoVE, JoVE, 2021, ⟨10.3791/61660⟩ |
| ISSN: | 1940-087X |
| DOI: | 10.3791/61660⟩ |
| Popis: | International audience; Summary:This paper details the fabrication protocol of microfluidic chips developed for on-chip protein crystallization with the dialysis method and in situ X-ray diffraction experiments. The microfabrication process makes it possible to integrate a semipermeable regenerated cellulose dialysis membrane with any molecular weight cut-off, between two layers of the chip.Abstract:This protocol describes the manufacturing of reproducible and inexpensive microfluidic devices covering the whole pipeline for crystallizing proteins on-chip with the dialysis method and allowing in situ single-crystal or serial crystallography experiments at room temperature. The protocol details the fabrication process of the microchips, the manipulation of the on-chip crystallization experiments and the treatment of the in situ collected X-ray diffraction data for the structural elucidation of the protein sample. The main feature of this microfabrication procedure lies on the integration of a commercially available, semipermeable regenerated cellulose dialysis membrane in between two layers of the chip. The molecular weight cut-off of the embedded membrane varies depending on the molecular weight of the macromolecule and the precipitants. The device exploits the advantages of microfluidic technology, such as the use of minute volumes of samples ( |
| Databáze: | OpenAIRE |
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