Journal article · Preprint article
Systematic investigation of protein phase behavior with a microfluidic formulator
Department of Applied Physics, California Institute of Technology, MS 128-95, Pasadena, CA 91125
We demonstrated a microfluidic device for rapidly generating complex mixtures of 32 stock reagents in a 5-nl reactor. This “formulation chip” is fully automated and allows thousands of experiments to be performed in a single day with minimal reagent consumption. It was applied to systematically study the phase behavior of the protein xylanase over a large and complex chemical space.
For each chemical formulation that demonstrated a pronounced effect on solubility, the protein phase behavior was completely mapped in the chip, generating a set of empirical phase diagrams. This ab initio phase information was used to devise a rational crystallization screen that resulted in 72-fold improvement in successful crystallization hits compared with conventional sparse matrix screens.
This formulations tool allows a physics-based approach to protein crystallization that may prove useful in structural genomics efforts.
Language: | Undetermined |
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Publisher: | National Academy of Sciences |
Year: | 2004 |
Pages: | 14431-14436 |
ISSN: | 10916490 and 00278424 |
Types: | Journal article and Preprint article |
DOI: | 10.1073/pnas.0405847101 |