Protein-based conjugates and self-assembled nanostructures

Patent metadata

Field	Value
Publication number	US-9359457-B2
Application number	US-201113882035-A
Country	US
Kind code	B2
Filing date	Oct 26, 2011
Priority date	Oct 26, 2010
Publication date	Jun 7, 2016
Grant date	Jun 7, 2016

Abstract

The present disclosure provides protein-containing compositions, methods and uses thereof. A conjugate comprises a globular protein conjugated with a polymer that preserves the folded and functional structure of a protein. In some embodiments, a fusion protein comprises a globular protein conjugated with an elastin-mimic polymer (EMP). Also disclosed is an assembled solid-state or gel-state nanostructure comprising a plurality of conjugates, wherein each comprises a globular protein conjugated with a polymer that preserving the folded and functional structure of a protein. In certain embodiments, provided compositions and methods further comprise an additive.

First claim

We claim:

1. An assembled solid-state or gel-state nanostructure comprising: a plurality of conjugates, wherein each comprises a globular protein conjugated with a polymer that preserves the folded and functional structure of a protein and wherein the weight percentage of the globular protein in the assembled nanostructure is greater than 20%.

2. The assembled nanostructure of claim 1 , further comprising an additive.

3. An assembled solid-state or gel-state nanostructure comprising: a plurality of fusion proteins, wherein each comprises a globular protein conjugated with an elastin-mimic polymer (EMP) and wherein the weight percentage of the globular protein in the assembled nanostructure is greater than 20%.

4. A method of forming an assembled nanostructure, comprising:
providing a plurality of conjugates in an aqueous solution, wherein each of the conjugates comprises a globular protein conjugated with a polymer that preserves the folded and functional structure of the protein;
removing the aqueous solution so that the conjugates self-assemble to form a solid-state or gel-state nanostructure, wherein the weight percentage of the globular protein in the assembled nanostructure is greater than 20%.

5. The method of claim 4 , further comprising a step of adding an additive to the aqueous solution before removing the aqueous solution.

6. The assembled nanostructure of claim 1 , wherein the assembled nanostructure has a lamellar structure, a perforated lamellar structure, and/or a hexagonally packed structure.

7. The assembled nanostructure of claim 1 , wherein the globular protein is an enzyme.

8. The assembled nanostructure of claim 1 , wherein the polymer is a synthetic polymer.

9. The assembled nanostructure of claim 1 , wherein the globular protein is covalently conjugated to the polymer.

10. The assembled nanostructure of claim 1 , wherein the polymer has a transition temperature greater than 0° C.

11. The assembled nanostructure of claim 6 , wherein the polymer has a transition temperature less than 80° C.

12. The assembled nanostructure of claim 1 , wherein the polymer has a transition temperature in a range of 25° C. to 70° C.

13. The assembled nanostructure of claim 1 , wherein the polymer has a molecular weight greater than 5 kDa.

14. The assembled nanostructure of claim 1 , wherein the polymer has a molecular weight in a range of 5 kDa to 100 kDa.

15. The assembled nanostructure of claim 8 , wherein the polymer is selected from the group consisting of poly(N-isopropylacrylamide), poly(3-[N-(2-methacroyloyethyl)-N,N-dimethylammonio]propane-sulfonate), poly(hydroxypropyl acrylate), poly(ethylene glycol methyl ether acrylate-b-methoxyethoxy ethyl acrylate) and any combination thereof.

16. The assembled nanostructure of claim 1 , wherein the globular protein is a fluorescent protein.

17. The assembled nanostructure of claim 1 , wherein the molecular weight of the globular protein is greater than 10 kDa.

18. The assembled nanostructure of claim 3 , wherein the EMP comprises the repeat unit of Val Pro Gly Xaa Gly (SEQ ID No:1), wherein Xaa is selected from the group consisting of Val, Leu, Ile, Ala and Gly.

Assignees

• Olsen Bradley D
• Thomas Carla S
• Massachusetts Inst Technology

Inventors

• Olsen Bradley D
• Thomas Carla S

Classifications

• B82Y5/00 Primary

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• C12N9/96

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• C08F20/56 Primary

  Acrylamide; Methacrylamide · CPC title

• A61K47/48176

  Human Necessities · mapped topic

• A61K47/58

  obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin · CPC title