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Systematic investigation of protein phase behavior with a microfluidic formulator

Hansen, Carl L. and Sommer, Morten O. A. and Quake, Stephen R. (2004) Systematic investigation of protein phase behavior with a microfluidic formulator. Proceedings of the National Academy of Sciences of the United States of America, 101 (40). pp. 14431-14436. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:HANpnas04

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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.


Item Type:Article
Additional Information:© 2004 by The National Academy of Sciences of the USA Communicated by Robert H. Austin, Princeton University, Princeton, NJ, August 16, 2004 (received for review February 9, 2004) Published online before print September 27, 2004, 10.1073/pnas.0405847101 [From The Cover] We thank Sebastian Maerkl and James Berger for helpful conversations and insights. This work was supported by the National Science Foundation, the Defense Advanced Research Planning Agency, and the Army Research Office. C.L.H. was partially funded by the Natural Sciences and Engineering Research Council (Canada) (Julie Payette Fellowship).
Subject Keywords:positive displacement crossinjection
Record Number:CaltechAUTHORS:HANpnas04
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:HANpnas04
Alternative URL:http://dx.doi.org/10.1073/pnas.0405847101
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:686
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:14 Sep 2005
Last Modified:14 Nov 2014 19:18

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