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首页> 外文期刊>Crystal growth & design >Microbatch Mixing: 'Shaken not Stirred', a Method for Macromolecular Microcrystal Production for Serial Crystallography
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Microbatch Mixing: 'Shaken not Stirred', a Method for Macromolecular Microcrystal Production for Serial Crystallography

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摘要

The advances of serial crystallography techniques at synchrotron and X-ray free electron laser facilities have made possible the acquisition of useable data sets to determine 3-dimensional structures of macromolecules from micro- to nanosized crystals. In addition, the same technological hallmarks have contributed significantly to the field of time resolved crystallography. However, the production of usable crystalline slurries for serial crystallographic experiments has been one of the limiting factors and contributes to an alternative sample "bottleneck" in crystal growth. In this study, we propose a method: labeled microbatch mixing (MBM), which has the capability to produce large quantities of microcrystals crystallographic experiments. This is shown to be successful for producing lysozyme, carbonic anhydrase, and adeno-associated virus crystals. MBM takes advantage of secondary nucleation induced by mixing via the application of steady agitation during the crystallization process. This leads to excessive nucleation, resulting in large quantities of well-diffracting microcrystals. MBM therefore presents a method that can potentially be applied to a range of macromolecules and a possible simple protocol to produce microcrystals for serial crystallographic experiments.

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