Back to results
US 2026/0265318 A1Published application
Method of producing soluble microdystrophin protein and uses of the same
AssigneeUltragenyx Pharmaceutical Inc.
InventorsJaclyn Cadaoas, James Lynden Bodnar, Mark Fox
Life of this patent
Estimating the term…
Abstract
The present disclosure provides novel methods of producing soluble human microdystrophin proteins and uses of the same including for in vitro detection assays.
Claims35
A method of producing a recombinant human microdystrophin (rhMD) protein in solution, the rhMD protein comprising:
- an N-terminal actin binding domain (NTD);
- one to five Spectrin-Like Repeat (R) domains selected from R1 domain, R2 domain, R3 domain, R16 domain, R17 domain, R22 domain, R23 domain, and R24 domain;
- and a beta-dystroglycan binding domain;
- the method comprising:
- (a) transfecting a host cell culture with a nucleic acid encoding the rhMD protein;
- (b) expressing the rhMD protein in the host cell culture;
- (c) generating a cell lysate by incubating the host cell culture with a lysis buffer;
- (d) centrifuging the cell lysate to generate a cell membrane fraction, said cell membrane fraction comprising plasma membranes of the cell culture;
- and
- (e) contacting the cell membrane fraction with a membrane extraction buffer comprising an amount of an anionic detergent to release the rhMD protein from the cell membrane fraction.
7 more elements
2-9. (canceled)
11-12. (canceled)
14-31. (canceled)
(canceled)
(canceled)
38-39. (canceled)
41-48. (canceled)
(canceled)
54-57. (canceled)
59-62. (canceled)
-67. (canceled)
70-82. (canceled)
85-91. (canceled)
93-100. (canceled)
(canceled)
Dependencies are inferred from the claim text.