Method for preparing protein cage, and in situ method for preparing hydrophobic additive-supported core-shell structured polymer-protein particles

Patent metadata

Field	Value
Publication number	US-9757342-B2
Application number	US-201314778906-A
Country	US
Kind code	B2
Filing date	Oct 11, 2013
Priority date	Mar 22, 2013
Publication date	Sep 12, 2017
Grant date	Sep 12, 2017

Abstract

The present invention relates to a method for preparing a protein cage which comprises: a 1 st step of preparing an amphiphilic polymer comprising a 1 st hydrophobic polymer and a 1 st hydrophilic functional group; a 2 nd step of preparing a hydrophilic protein comprising a 2 nd functional group binding to the 1 st functional group; a 3 rd step of forming an amphiphilic polymer-protein hybrid by the binding of the 1 st functional group and the 2 nd functional group, and forming core-shell structured particles comprising a protein shell and an amphiphilic polymer core by the self-assembly of the amphiphilic polymer in a hydrophilic solvent; and a fourth step of removing some or all of the hydrophobic polymer of the core part from the core-shell structured particles.

First claim

The invention claimed is:

1. A method for preparing a protein cage which comprises:
a 1 st step of preparing an amphiphilic polymer comprising a 1 st hydrophobic polymer and a 1 st hydrophilic functional group;
a 2 nd step of preparing a hydrophilic protein comprising a 2 nd functional group binding to the 1 st functional group;
a 3 rd step of forming an amphiphilic polymer-protein hybrid by the binding of the 1 st functional group and the 2 nd functional group, and forming core-shell structured particles comprising a protein shell and an amphiphilic polymer core by the self-assembly of the amphiphilic polymer in a hydrophilic solvent; and
a 4 th step of removing some or all of the hydrophobic polymer of the core part from the core-shell structured particles.

2. The method of claim 1 , wherein the protein retains its activity by maintaining its tertiary structure.

3. The method of claim 1 , wherein the protein cage comprises one or at least two types of proteins.

4. The method of claim 1 , wherein the method further comprises a step of forming bindings between the shell-forming proteins by adding a cross-linking agent to the core-shell structured particles which are formed in the 3 rd step.

5. The method of claim 1 , wherein a 2 nd hydrophobic polymer not having the 1 st functional group is further added during the 3 rd step.

6. The method of claim 1 , wherein an additive is loaded in the protein cage.

7. The method of claim 1 , wherein the additive is added in the 3 rd step to be included in the core part during the self-assembly or is injected into the protein cage formed in the fourth step.

8. The method of claim 1 , wherein removal of some or all of the hydrophobic polymer of the core part from the core-shell structured particles in the fourth step is performed by the introduction of (i) a competitor compound for the binding between the 1 st and the 2 nd functional groups, or (ii) a compound that hydrolyzes the polymer moiety in the amphiphilic polymer-protein hybrid.

Assignees

• Pusan Nat Univ Industry—Univ Coop Found
• Korea Res Inst Chemical Tech
• Gachon Univ Of Industry—Academic Coop Found
• Pusan Nat Univ Industry-Univ Coop Found
• Gachon Univ Of Industry-Academic Coop Found

Inventors

• Paik Hyun Jong
• Song Jae Kwang
• An Seong Soo
• Lee Chae Yeon
• Jeong Jong Hwa
• Kadir Mohammad Abdul
• Lee Tae Heon

Classifications

• A61K8/66

  Enzymes · CPC title

• A61K8/0204 Primary

  Specific forms not provided for by any of groups A61K8/0208 - A61K8/14 · CPC title

• A61Q19/00

  Preparations for care of the skin · CPC title

• A61K9/5138

  obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates · CPC title

• C07K2319/21

  containing a His-tag · CPC title