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Capillary technologies |
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Over the last 5 to 7 years, small drop crystallisation robotics has revolutionised crystallisation for protein structure determination. A description of the state of the art crystallisation facilities in Europe can be found in the WP5 description. A key aim of the developmental work outlined here is to overcome the reality that obtaining crystals of sufficient quality for structural determination is currently a bottleneck in the pipeline process (Berry et al., Acta Cryst D62:1137, 2006). We focus on two well-defined aspects of the crystallisation process where we expect to have a positive impact on the effectiveness of the current infrastructure, namely the use of counter diffusion capillary crystallisation and the use of in situ X-ray diffraction measurements of crystallisation experiments.
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Capillaries. The aim is to change the format of capillary crystallisation and develop a low-cost technology that is suitable for automation and subsequent direct X-ray diffraction analysis. The physics of the counter diffusion process in capillaries is different to the usual vapour diffusion method making the experiment more robust and less sensitive to protein concentration. The capillary format is also ideal for direct diffraction experiments without further manipulation of fragile crystals (e.g. for membrane proteins and protein complexes). This links directly to the second major development:
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In situ X-ray diffraction screening of crystals. With this technology the diffraction quality of crystals can be assessed immediately, eliminating the error-prone steps of crystal manipulation and freezing. Optimisation efforts can be focussed on the best diffracting crystals and salt crystals can be identified and discarded at an early stage. This will significantly shorten the optimisation loop for growing diffraction quality crystals.
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