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Nanoliter-scale protein crystallization and screening with a microfluidic droplet robot.

ABSTRACT: Large-scale screening of hundreds or even thousands of crystallization conditions while with low sample consumption is in urgent need, in current structural biology research. Here we describe a fully-automated droplet robot for nanoliter-scale crystallization screening that combines the advantages of both automated robotics technique for protein crystallization screening and the droplet-based microfluidic technique. A semi-contact dispensing method was developed to achieve flexible, programmable and reliable liquid-handling operations for nanoliter-scale protein crystallization experiments. We applied the droplet robot in large-scale screening of crystallization conditions of five soluble proteins and one membrane protein with 35-96 different crystallization conditions, study of volume effects on protein crystallization, and determination of phase diagrams of two proteins. The volume for each droplet reactor is only ca. 4-8?nL. The protein consumption significantly reduces 50-500 fold compared with current crystallization stations.


PROVIDER: S-EPMC5154416 | BioStudies | 2014-01-01T00:00:00Z

REPOSITORIES: biostudies

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