Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins.

Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Research Abstract Details 

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Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Abstract Text:

Liang Li,Debarshi Mustafi,Qiang Fu,Valentina Tereshko,Delai L Chen,Joshua D Tice,Rustem F Ismagilov,

High-throughput screening and optimization experiments are critical to a number of fields, including chemistry and structural and molecular biology. The separation of these two steps may introduce false negatives and a time delay between initial screening and subsequent optimization. Although a hybrid method combining both steps may address these problems, miniaturization is required to minimize sample consumption. This article reports a "hybrid" droplet-based microfluidic approach that combines the steps of screening and optimization into one simple experiment and uses nanoliter-sized plugs to minimize sample consumption. Many distinct reagents were sequentially introduced as approximately 140-nl plugs into a microfluidic device and combined with a substrate and a diluting buffer. Tests were conducted in approximately 10-nl plugs containing different concentrations of a reagent. Methods were developed to form plugs of controlled concentrations, index concentrations, and incubate thousands of plugs inexpensively and without evaporation. To validate the hybrid method and demonstrate its applicability to challenging problems, crystallization of model membrane proteins and handling of solutions of detergents and viscous precipitants were demonstrated. By using 10 microl of protein solution, approximately 1,300 crystallization trials were set up within 20 min by one researcher. This method was compatible with growth, manipulation, and extraction of high-quality crystals of membrane proteins, demonstrated by obtaining high-resolution diffraction images and solving a crystal structure. This robust method requires inexpensive equipment and supplies, should be especially suitable for use in individual laboratories, and could find applications in a number of areas that require chemical, biochemical, and biological screening and optimization.

Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Publishing Authors By Initials

L Li,D Mustafi,Q Fu,V Tereshko,DL Chen,JD Tice,RF Ismagilov,

For similar investigative techniques: chemistry, analytical: crystallography: x-ray diffraction research abstracts see: investigative techniques: chemistry, analytical: crystallography: x-ray diffraction research

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Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Proceedings of the National Academy of Sciences of

VOLUME: 103

Page Numbers: 19243-8

Journal Abbreviation: Proc. Natl. Acad. Sci. U.S.A.

ISSN: 0027-8424

DAY: 11

MONTH: 12

YEAR: 2006

Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 7505876

Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Keywords Mesh Terms:

KEYWORDS: X-Ray Diffraction

MESH TERMS: methods

Chemical & Substance for Abstract: Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins. Information

Substance Name: Membrane Proteins

Registry Number: 0

Grant and Affiliation Information for Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins.

AFFILIATION: Department of Chemistry and Institute for Biophysical Dynamics, University of Chicago, Chicago, IL 60637, USA.

Country: United States

AGENCY: United States NIGMS

GRANT: Y1-GM-1104

ACRONYM: GM

MEDLINETA: Proc Natl Acad Sci U S A

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