Microfluidic protein crystallography
First Claim
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1. A method of crystallization comprising:
- utilizing a microfluidic formulator device to generate a solubility fingerprint of a crystallization target over a range of conditions;
utilizing the microfluidic formulator to map phase space around precipitating conditions of the solubility fingerprint resulting in precipitation of the crystallization target and soluble conditions of the solubility fingerprint where the crystallization target precipitate dissolves; and
determining a hysteresis between precipitating and soluble phase spaces.
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Abstract
The use of microfluidic structures enables high throughput screening of protein crystallization. In one embodiment, an integrated combinatoric mixing chip allows for precise metering of reagents to rapidly create a large number of potential crystallization conditions, with possible crystal formations observed on chip. In an alternative embodiment, the microfluidic structures may be utilized to explore phase space conditions of a particular protein crystallizing agent combination, thereby identifying promising conditions and allowing for subsequent focused attempts to obtain crystal growth.
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Citations
30 Claims
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1. A method of crystallization comprising:
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utilizing a microfluidic formulator device to generate a solubility fingerprint of a crystallization target over a range of conditions; utilizing the microfluidic formulator to map phase space around precipitating conditions of the solubility fingerprint resulting in precipitation of the crystallization target and soluble conditions of the solubility fingerprint where the crystallization target precipitate dissolves; and determining a hysteresis between precipitating and soluble phase spaces. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A crystallization method comprising:
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empirically determining a solubility curve for a crystallization target mixed with a precipitant utilizing a microfluidic device; and mixing the crystallization target with the precipitant at a ratio that places a final concentration of the crystallization target and the precipitant on a boundary of the solubility curve. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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Specification