IP Library Granted Patent US 10,526,395
Granted Patent B2
US 10,526,395 · App. 15/763,052 · Granted Jan 7, 2020

Detergent-protein composition comprising lyophilized detergent-solubilized protein

Inventors: Michael F. Brown (Tucson, AZ); Suchithranga Perera (Tucson, AZ); Udeep Chawla (Tucson, AZ)
Assignee: ARIZONA BOARD OF REGENTS ON BEHALF OF THE UNIVERSITY OF ARIZONA
C07K14/723
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Quick Facts
Patent No.
US 10,526,395
App. No.
15/763,052
Granted
Jan 7, 2020
Kind
B2
Abstract

A method for generating detergent-solubilized G-protein coupled receptors (GPCRs) in powdered form. The powdered GPCRs is storable at temperatures of −20° C. or lower and for durations of 365 days or longer without substantial loss of functionality of the GPCRs after rehydration as compared to a pre-frozen state. The method can maintain minimal detergent to protein ratio in molar concentrations. The method can further generate GPCRs with specific water or deuterium content as required in certain experiments, such as mass spectrometry, NMR spectroscopy, or quasi-elastic neutron scattering (QENS).

Claims (7)

1. A detergent-protein composition comprising a lyophilized detergent-solubilized protein, said detergent-solubilized protein comprising an amphiphilic membrane protein and a detergent, wherein a molar ratio of the detergent to amphiphilic protein is at most about 30:1, wherein the amphiphilic membrane protein is not irreversibly denatured.

2. The composition of claim 1 , wherein the amphiphilic protein is a G-protein-coupled receptor (GPCR).

3. The composition of claim 1 , wherein the detergent is a nonionic, an anionic, a cationic or a zwitterionic detergent.

4. The composition of claim 1 , wherein the detergent-protein composition is rehydrated from a lyophilized state, wherein the amphiphilic protein regains its photochemical functionality and maintains its structure as compared to its unpreserved state.

5. The composition of claim 1 , wherein the functionality and structure of the amphiphilic protein is maintained in the lyophilized state.

6. The composition of claim 5 , wherein the composition is in a powdered form, wherein the powdered composition is capable of being stored at a temperature of at most about −20° C. and for a duration of at least about 365 days, wherein the powdered composition is capable of being rehydrated after being stored, wherein when the powdered composition is rehydrated, the amphiphilic protein regains its photochemical functionality and maintains its structure as compared to its unpreserved state.

7. A detergent-protein composition comprising a lyophilized detergent-solubilized protein, said detergent-solubilized protein comprising an amphiphilic membrane protein and a detergent, wherein the amphiphilic membrane protein is not irreversibly denatured.

Assignments (2)
CONFIRMATORY LICENSE Recorded May 10, 2018
From: UNIVERSITY OF ARIZONA
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 046125/0088 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 26, 2018
From: BROWN, MICHAEL F.; PERERA, SUCHITHRANGA; CHAWLA, UDEEP
To: ARIZONA BOARD OF REGENTS ON BEHALF OF THE UNIVERSITY OF ARIZONA
Reel/Frame 045353/0308 →
Continuity (3)
Provisional Application 62263295 · Dec 4, 2015
Provisional Application 62235335 · Sep 30, 2015
Related Publication 20180273606A1 · Sep 27, 2018