1. A chemical library to screen protein and protein-ligand crystallization using a versatile microfluidic platform
- Author
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Stéphane Veesler, Laurent Vuillard, Nadine Candoni, Romain Grossier, Gilles Ferry, Charline J. J. Gerard, Nathalie Ferte, Jean A. Boutin, Centre Interdisciplinaire de Nanoscience de Marseille (CINaM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherches Servier, and Centre de Recherches de Croissy
- Subjects
0301 basic medicine ,Materials science ,Microfluidics ,Nanotechnology ,010402 general chemistry ,01 natural sciences ,Chemical library ,law.invention ,03 medical and health sciences ,chemistry.chemical_compound ,law ,General Materials Science ,[SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering ,Crystallization ,Biological macromolecules ,Aqueous solution ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,General Chemistry ,Condensed Matter Physics ,Ligand (biochemistry) ,0104 chemical sciences ,030104 developmental biology ,chemistry ,Microfluidic ,[PHYS.COND.CM-MS]Physics [physics]/Condensed Matter [cond-mat]/Materials Science [cond-mat.mtrl-sci] ,Protein crystallization ,Co-crystallization ,Microfabrication ,Protein ligand - Abstract
International audience; Here, we describe a plug-and-play microfluidic platform, suitable for protein crystallization. The droplet factory is designed to generate hundreds of droplets as small as a few nanoliters (2 to 10nL) for screening and optimization of crystallization conditions. Commercially-available microfluidic junctions and tubing are combined to create the appropriate geometry. In addition, a " chemical library " is produced in tubing. The microfluidic geometry for a " crystallization agent-based chemical library " is validated by screening crystallization conditions of lysozyme. The microfluidic geometry for a " ligand-based chemical library " is also explored to co-crystallize the protein QR2 with a ligand, for the purposes of structure-based drug design. This platform mixes aqueous phases (containing the protein and the crystallization agent) and organic phases (containing the ligand), during the droplet generation and circulation without using any surfactant. The droplet composition is controlled by the respective flow-rates of the different solutions, and checked by measuring on-line absorbance. The low volumes involved in the crystallization trials, the speed of execution and the absence of a microfabrication stage make our platform a cheap, easy-to-use and versatile tool for crystallization studies.
- Published
- 2018