Abstract
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.
Original language | English |
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Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 101 |
Issue number | 40 |
Pages (from-to) | 14431-14436 |
Number of pages | 6 |
ISSN | 0027-8424 |
DOIs | |
Publication status | Published - 2004 |
Externally published | Yes |
Keywords
- Biophysical Phenomena
- Biophysics
- Chemical Precipitation
- Crystallization
- Endo-1,4-beta Xylanases
- Microfluidics
- Proteins
- Solubility
- Spectrophotometry
- Trichoderma
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