Compositions and methods for membrane protein detergent stability screen

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United States of America Patent

PATENT NO 9606126
APP PUB NO 20120108463A1
SERIAL NO

13376729

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Abstract

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The invention provides methods to assess protein stability and to obtain sizing information. In one aspect, the screen comprises a 94 detergent panel and a series of MWCO filtered microplates. A protein of interest is bound to an affinity matrix and aliquoted into a 96-well microplate. Wells containing the immobilized protein are washed in the new detergent and then eluted in the new detergent into a collection plate. Protein not stable in the new detergent is precipitated on the resin and not present in the elutions. Half of the elution is passed through a high (i.e., 300 kDa) MWCO microplate and the other half through a low (i.e., 100 kDa) MWCO microplate. Elutions from the microplates are spotted on a nitrocellulose membrane, visualized by Western analysis (or by some other method), and quantified. The high MWCO provides stability readout and the ratio of low/high kDa provides sizing information.

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Patent Owner(s)

Patent OwnerAddress
UNIVERSITY OF VIRGINIA PATENT FOUNDATION250 WEST MAIN STREET SUITE 300 CHARLOTTESVILLE VA 22902

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Inventor(s)

Inventor Name Address # of filed Patents Total Citations
Vergis, James M Gaithersburg, US 2 0
Wiener, Michael C Charlottesville, US 4 3

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