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Automated microfluidic sample‐preparation platform for high‐throughput structural investigation of proteins by small‐angle X‐ray scattering
Author(s) -
Lafleur Josiane P.,
Snakenborg Detlef,
Nielsen Søren S.,
Møller Magda,
Toft Katrine N.,
Menzel Andreas,
Jacobsen Jes K.,
Vestergaard Bente,
Arleth Lise,
Kutter Jörg P.
Publication year - 2011
Publication title -
journal of applied crystallography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.429
H-Index - 162
ISSN - 1600-5767
DOI - 10.1107/s0021889811030068
Subject(s) - small angle x ray scattering , microfluidics , absorbance , fluidics , throughput , mixing (physics) , scattering , sample (material) , materials science , sample preparation , analytical chemistry (journal) , x ray , chemistry , chromatography , nanotechnology , optics , computer science , physics , engineering , telecommunications , quantum mechanics , aerospace engineering , wireless
A new microfluidic sample‐preparation system is presented for the structural investigation of proteins using small‐angle X‐ray scattering (SAXS) at synchrotrons. The system includes hardware and software features for precise fluidic control, sample mixing by diffusion, automated X‐ray exposure control, UV absorbance measurements and automated data analysis. As little as 15 µl of sample is required to perform a complete analysis cycle, including sample mixing, SAXS measurement, continuous UV absorbance measurements, and cleaning of the channels and X‐ray cell with buffer. The complete analysis cycle can be performed in less than 3 min. Bovine serum albumin was used as a model protein to characterize the mixing efficiency and sample consumption of the system. The N2 fragment of an adaptor protein (p120‐RasGAP) was used to demonstrate how the device can be used to survey the structural space of a protein by screening a wide set of conditions using high‐throughput techniques.