Microfluidic Network for Droplets: Store, then Create
- 技术优势
- ·Useful forprotein crystallization or other applications requiring formulation and storageof many small samples of nano-liter volumes.· Eliminatescross contamination between samples.· Cheapermethod to test for the optimal concentration of protein.· Crystalscan be small which increases the potential for growing crystals in the firstplace.· Thepresent method avoids the roughly tenfold increase in crystal mosaicitytypically encountered during cryoprotection and eliminates the need to searchfor cryoprotectant conditions.
- 详细技术说明
- Microfluidic Network for Droplets: Store, then Create
- *Abstract
-
Thepresent invention relates to a method for simultaneous generation and storageof isolated droplets of aqueous solutions. To date, existing methods used todetermine the optimal concentrations for achieving protein crystallization haveincluded mixing macro-portions of multiple aqueous solutions. This mixingoccurs in testing trays called crystallization trays which have severalchambers and contain the mixtures of proteins at various concentrations. Thedisadvantage of the current method is that often due to the macro sized volumeof these testing trays, it becomes cost prohibitive to test out multiplesolution concentrations at one time. Additionally, in conventional proteinX-ray crystallography, a large crystal is required in order to obtain acomplete data set.
Ourinvention is based on an emulsion-based serial crystallographic technology inwhich nano-litre sized droplets of protein solutions and encapsulated in oiland stabilized by surfactant. Serial crystallography takes the oppositeapproach to traditional X-ray crystallography techniques of developing onelarge crystal. In this instance, a complete diffraction set is assembled from alarge number of individual diffraction frames acquired from small singleunoriented crystals that are not cryoprotected. Our microfluidic device based containsmultiple sequences of capillary valves and storage chambers that simply androbustly generate and store aqueous solutions in chambers that are isolatedfrom one another by oil. The microfluidic chip is able to generate drops in aserial fashion. The device has the capability of allowing different chemicals tobe added without mixing, or alternatively, will allow for chemicals to be addedto the mix in pre-determined quantities set by the geometry of the device.
Thisemulsion-based serial crystallographic technology, in which nano-litre sized dropletsof protein solution are encapsulated in oil and stabilized by surfactant worksby allowing the first crystal in a drop to be nucleated and this small volumethen generates a negative feedback mechanism that lowers the supersaturation. Thismechanism is exploited to produce one crystal per drop. Diffraction data aremeasured, one crystal at a time, from a series of room temperature crystalsstored on an X-ray semi-transparent microfluidic chip, and a 93% complete dataset is obtained by merging single diffraction frames taken from differentunoriented crystals. This process is cheaper and provides better qualitycrystals for protein crystallization over the current methods.
- *IP Issue Date
- None
- *IP Type
- Other Patent
- 国家
- United States
- 申请号码
- PCT/US2018/013877
- 国家/地区
- 美国
