By Patrick W. Cooley, David Silva, David B. Wallace, MicroFab Technologies, Inc., Stephen Sprang, Dianne Decamp, University of Montana
X-ray crystallography is used routinely by scientists to obtain the three dimensional structure of a biological molecule of interest.Such information can be used to determine how a pharmaceutical interacts with a protein target and what changes might improve functionality. However, the crystallization of macromolecules still remains a serious hindrance in structural determination despite impressive advances in screening methods and technologies.
CrystalJet is a prototype inkjet based platform to perform small fluid volume high-throughput protein crystallization screening to identify parameters for obtaining diffraction quality crystals. CrystalJet provides decreased protein consumption by using smaller screening volumes, thus maximizing the sampling of crystal growth conditions. The CrystalJet screening platform will operate in microbatch and vapor diffusion modes to offer experimental flexibility. The system utilizes aspirate and dispense technology to perform the aspiration of the crystallization precipitant solutions with piezoelectric dispensing at highly repeatable picoliter (100) to nanoliter volumes, the latter via drop accretion. A separate disposable inkjet dispensing device is used to deliver the protein solution alleviating cross contamination issues. The platform includes a plate sealer and environmental control to maintain temperature and humidity.
A series of experiments validated the potential use of CrystalJet for the screening of macromolecule crystallization conditions using ultra-small drops of protein and crystallization solution. This inkjet-based system can be used for protein crystallization screening to maximize the conservation of protein samples.
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