Cyclic olefin homopolymer-based microfluidics for protein crystallization and in situ X-ray diffraction

Article Article scientifique Article Cyclic olefin homopolymer-based microfluidics for protein crystallization and in situ X-ray diffraction eng 2009-08-06 2009-08-06 author Josserand Soheila 555363 author Petty Tom J. author De Almeida Victor 554550 author Nishimura Taisuke 636978 author Joly Jacques author Ferrer Jean-Luc author Halazonetis Thanos 641033 Département de biologie moléculaire et cellulaire (MOCEL) 913-920 10.1107/S0907444909021489 19690369 0907-4449 PMC2733880 https://scripts.iucr.org/cgi-bin/paper?S0907444909021489 <p>Microfluidics is a promising technology for the rapid identification of protein crystallization conditions. However, most of the existing systems utilize silicone elastomers as the chip material which, despite its many benefits, is highly permeable to water vapour. This limits the time available for protein crystallization to less than a week. Here, the use of a cyclic olefin homopolymer-based microfluidics system for protein crystallization and in situ X-ray diffraction is described. Liquid handling in this system is performed in 2 mm thin transparent cards which contain 500 chambers, each with a volume of 320 nl. Microbatch, vapour-diffusion and free-interface diffusion protocols for protein crystallization were implemented and crystals were obtained of a number of proteins, including chicken lysozyme, bovine trypsin, a human p53 protein containing both the DNA-binding and oligomerization domains bound to DNA and a functionally important domain of Arabidopsis Morpheus' molecule 1 (MOM1). The latter two polypeptides have not been crystallized previously. For X-ray diffraction analysis, either the cards were opened to allow mounting of the crystals on loops or the crystals were exposed to X-rays in situ. For lysozyme, an entire X-ray diffraction data set at 1.5 A resolution was collected without removing the crystal from the card. Thus, cyclic olefin homopolymer-based microfluidics systems have the potential to further automate protein crystallization and structural genomics efforts.</p> Dewey 540 Dewey 580 Dewey 570 39 Département de biochimie 144 43 Département de biologie moléculaire 161 45 Département des sciences végétales 163 false Acta crystallographica. Section D, Biological crystallography 65 9 02f2d1ec-bf41-4770-94ed-91168652e1dc Article (Published version) Public Article-scientifique-from-unpaywall.pdf 994783 application/pdf 0b6001425ecc4f8c62578a0b67ef50ee4565eb60c3411b35ba330a3e6606406b openAccess Faculté des sciences / Section de chimie et biochimie / Département de biochimie Faculté des sciences / Section de biologie / Département de biologie moléculaire Faculté des sciences / Section de biologie / Département des sciences végétales

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Cyclic olefin homopolymer-based microfluidics for protein crystallization and in situ X-ray diffraction	Acta crystallographica. Section D, Biological crystallography			2009	57	63

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De Almeida, Victor