Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis.
Microfluidic devices were designed to perform on micromoles of biological macromolecules and viruses the search and the optimization of crystallization conditions by counter-diffusion, as well as the on-chip analysis of crystals by X-ray diffraction. Chips composed of microchannels were fabricated in poly-dimethylsiloxane (PDMS), poly-methyl-methacrylate (PMMA) and cyclo-olefin-copolymer (COC) by three distinct methods, namely replica casting, laser ablation and hot embossing. The geometry of the channels was chosen to ensure that crystallization occurs in a convection-free environment. The transparency of the materials is compatible with crystal growth monitoring by optical microscopy. The quality of the protein 3D structures derived from on-chip crystal analysis by X-ray diffraction using a synchrotron radiation was used to identify the most appropriate polymers. Altogether the results demonstrate that for a novel biomolecule, all steps from the initial search of crystallization conditions to X-ray diffraction data collection for 3D structure determination can be performed in a single chip.
Architecture et réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084, Strasbourg, France.
Kaouthar
K
Dhouib
Chantal
C
Khan Malek
Wilhelm
W
Pfleging
Bernard
B
Gauthier-Manuel
Roland
R
Duffait
Gaël
G
Thuillier
Rosaria
R
Ferrigno
Lilian
L
Jacquamet
Jeremy
J
Ohana
Jean-Luc
JL
Ferrer
Anne
A
Théobald-Dietrich
Richard
R
Giegé
Bernard
B
Lorber
Claude
C
Sauter
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000295:instrumentation*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D046911:Macromolecular Substances / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D019904:Polymethyl Methacrylate / Q000737:chemistry
D004129:Dimethylpolysiloxanes; D046911:Macromolecular Substances; C013830:baysilon; D019904:Polymethyl Methacrylate