Abstract
The Microcapillary Protein Crystallization System (MPCS) embodies a new semi-automated plug-based crystallization technology which enables nanolitre-volume screening of crystallization conditions in a plasticware format that allows crystals to be easily removed for traditional cryoprotection and X-ray diffraction data collection. Protein crystals grown in these plastic devices can be directly subjected to in situ X-ray diffraction studies. The MPCS integrates the formulation of crystallization cocktails with the preparation of the crystallization experiments. Within microfluidic Teflon tubing or the microfluidic circuitry of a plastic CrystalCard, approximately 10-20 nl volume droplets are generated, each representing a microbatch-style crystallization experiment with a different chemical composition. The entire protein sample is utilized in crystallization experiments. Sparse-matrix screening and chemical gradient screening can be combined in one comprehensive ;hybrid' crystallization trial. The technology lends itself well to optimization by high-granularity gradient screening using optimization reagents such as precipitation agents, ligands or cryoprotectants.
Highlights
The field of structural biology is generating technologies that increase throughput and efficiency each year
The underlying technology used to perform microfluidic nanovolume crystallization discussed here was developed at the University of Chicago
Performing sparse-matrix and gradient screening in the same trial allows the Microcapillary Protein Crystallization System (MPCS) to scan a substantial portion of crystallization phase space, generating 20–40 individual crystallization experiments at different concentrations from each pre-formed crystallant plug
Summary
The field of structural biology is generating technologies that increase throughput and efficiency each year. Such advances have initiated a goal to progress from gene to three-dimensional structure in 3 d. The CrystalCards are manufactured with materials that enable a balance between X-ray transmission, optical clarity, moldability, chemical resistance and surface energy. This system produces diffraction-ready crystals by allowing crystal extraction from the CrystalCard or in situ X-ray. D64, 1116–1122 research papers diffraction and structure solution using diffraction data obtained from samples within the CrystalCard
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