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Accueil du site > Publications > Archives 2004-2010 > 2007 > Using Microfluidics to Decouple Nucleation and Growth of Protein Crystals

Using Microfluidics to Decouple Nucleation and Growth of Protein Crystals

J.-U. Shim, G. Cristobal, D. R. Link, T. Thorsen, S. Fraden

10 janvier 2006

A high-throughput, low-volume microfluidic device has been designed to decouple the physical processes of protein crystal nucleation and growth. This device, called the Phase Chip, is constructed out of poly(dimethylsiloxane) (PDMS) elastomer. One of the Phase Chip ?s innovations is to exploit surface tension forces to guide each drop to a storage chamber. We demonstrate that nanoliter water-in-oil drops of protein solutions can be rapidly stored in individual wells, thereby allowing the screening of 1000 conditions while consuming a total of only 10 µg of protein on a 20 cm2 chip. Another significant advance over current microfluidic devices is that each well is in contact with a reservoir via a dialysis membrane through which only water and other low-molecular-weight organic solvents can pass, but not salt, polymer, or protein. This enables the concentration of all solutes in a solution to be reversibly, rapidly, and precisely varied in contrast to current methods, such as the free interface diffusion or sitting drop methods, which are irreversible. The Phase Chip operates by first optimizing conditions for nucleation by using dialysis to supersaturate the protein solution, which leads to nucleation of many small crystals. Next, conditions are optimized for crystal growth by using dialysis to reduce the protein and precipitant concentrations, which leads small crystals to dissolve while simultaneously causing only the largest ones to grow, ultimately resulting in the transformation of many small, unusable crystals into a few large ones.

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