X-ray Compatible Microfluidic platforms for screening, Crystallization and De Novo Structure Determination of Proteins
- UIUC
We present here an X-ray compatible, array based microfluidic chip that functions as a complete crystallization platform. It is capable of screening multiple crystallization conditions using minimal amount of protein solution per trial and facilitates de novo structure determination by collecting and merging X-ray data from multiple crystals grown on-chip. Our approach bypasses issues associated with the need to grow large, high quality crystals, crystal damage during handling, cryocooling, and radiation exposure and enables in situ data collection at biologically relevant temperatures. To validate the method, we determined the structure of PhnA to 2.30{angstrom} resolution solely using data collected on-chip. Here we have presented a microfluidic platform that allows for screening of crystallization conditions, together with on-chip X-ray analysis of the crystals formed. Not only do these chips eliminate crystal handling, they also allow for room temperature data collection from multiple small crystals, as opposed to collecting data from single large crystals, which requires cryocooling to avoid radiation damage. Efforts on extending the capabilities of this platform to perform a variety of other studies on different novel protein targets are currently underway.
- Research Organization:
- Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
- Sponsoring Organization:
- NIH
- OSTI ID:
- 1031923
- Country of Publication:
- United States
- Language:
- ENGLISH
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X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection
X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection