en
Scientific article
Open access
English

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

Published inActa crystallographica. Section D, Biological crystallography, vol. 65, no. 9, p. 913-920
Publication date2009-08-06
First online date2009-08-06
Abstract

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.

eng
Citation (ISO format)
JOSSERAND, Soheila et al. Cyclic olefin homopolymer-based microfluidics for protein crystallization and in situ X-ray diffraction. In: Acta crystallographica. Section D, Biological crystallography, 2009, vol. 65, n° 9, p. 913–920. doi: 10.1107/S0907444909021489
Main files (1)
Article (Published version)
accessLevelPublic
Identifiers
ISSN of the journal0907-4449
8views
1downloads

Technical informations

Creation01/03/2024 9:13:04 AM
First validation02/08/2024 7:52:35 AM
Update time02/08/2024 7:52:35 AM
Status update02/08/2024 7:52:35 AM
Last indexation02/08/2024 7:52:36 AM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack