Nanolipoprotein particles and related methods and systems for protein capture, solubilization, and/or purification
AssigneeLawrence Livermore National Security, LLC
InventorsBrett A. Chromy, Paul Henderson, Paul D. Hoeprich, Jr.
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Abstract
Provided herein are methods and systems for assembling, solubilizing and/or purifying a membrane associated protein in a nanolipoprotein particle, which comprise a temperature transition cycle performed in presence of a detergent, wherein during the temperature transition cycle the nanolipoprotein components are brought to a temperature above and below the gel to liquid crystalling transition temperature of the membrane forming lipid of the nanolipoprotein particle.
Claims6
A method for assembling a membrane associated protein, a scaffold protein, and a membrane forming lipid into a nanolipoprotein particle, the membrane forming lipid having a membrane forming lipid gel-crystalline transition temperature, the method comprising
- contacting the membrane associated protein with the scaffold protein and the membrane forming lipid to provide an admixture;
- and
- subjecting the admixture to multiple temperature transition cycles in presence of a detergent, for a time and under condition to allow assembly of the nanolipoprotein particle, the temperature transition cycle comprising:
- a temperature increase step wherein the admixture is brought to a temperature above said membrane forming lipid gel crystalline transition temperature and
- a temperature decrease step wherein the admixture is brought to a temperature below said membrane forming lipid gel crystalline transition temperature.
Dependencies are inferred from the claim text.