Several aspects of health and medicinal research depend on the knowledge of macromolecular structures, the determination of which depends on a supply of high-quality crystals. Conditions for growing high-quality crystals are usually determined by expensive trial-and-error research. The proposed innovation is a lithography-fabricated system for automated, kinetically controlled growth of biomolecule crystals for crystallography. It is also useful in crystallization process development applications. An array of crystallization chambers, each with a volume of a few microlitres is served by an array of pumps whose feed rates are controlled in the sub-nL/s range by a time-of-flight flow meter. The pumps, one each for buffer, biomolecule solution and precipitant solution, are commanded by control algorithms that use image data. The individual concepts were demonstrated in Phase I research, namely, the ability to deliver a few microlitres of solution to a microreactor chamber over several hours and to grow crystals in such a chamber, the ability to electronically measure sub nL/s flow rates with a meter than has no moving parts, and the identification of a very inexpensive digital imaging and analysis method. The Phase II research specific aims are to (1) integrate these novel components into a single automated crystallization system to be tested for the coordination of all functions through a single control computer; (2) on the basis of (1) design and fabricate by lithography an integrated 4-chamber array including optics, microfluidics and electronics; and (3) test this array against real crystal-growth problems and refine image-based control algorithms. This technology uses automated kinetic control of the liquid-liquid crystallization method as a means of canvassing a continuum of chemical conditions rather than using massively paralleled discreet screening to enhance throughput in this rather expensive process. The resulting product, a crystallizer microarray (""""""""CRYMA""""""""), will be marketed to the world's large pharmaceutical firms, university laboratories with structural biology units and governmental space research agencies - a market with which SHOT, Inc. is familiar. Its implementation.
| Sugiyama, Masano; Sengupta, Shramik; Todd, Paul et al. (2008) Concentration control for protein crystallization via a continuously-fed crystallization chamber. Lab Chip 8:1398-401 |
