Fusion protein comprising diphtheria toxin non-toxic mutant CRM197 or fragment thereof

Patent metadata

Field	Value
Publication number	US-9757450-B2
Application number	US-201615248779-A
Country	US
Kind code	B2
Filing date	Aug 26, 2016
Priority date	Jun 1, 2011
Publication date	Sep 12, 2017
Grant date	Sep 12, 2017

Abstract

Provided in the present invention are a diphtheria toxin non-toxic mutant CRM197 or a fragment thereof as an adjuvant in a fusion protein and the use thereof to enhance the immunogenicity of a target protein fused therewith, for example, an HEV capsid protein, or an influenza virus M2 protein or an immunogenic fragment thereof. Also provided is a method for enhancing the immunogenicity of a target protein, comprising the fusion expression of the CRM197 or the fragment thereof with the target protein to form a fusion protein. Further provided is a fusion protein comprising the CRM197 or the fragment thereof and a target protein, the CRM197 or the fragment thereof enhancing the immunogenicity of the target protein. The present invention also provides an isolated nucleic acid encoding the fusion protein, a construct and a vector comprising said nucleic acid, and a host cell comprising the nucleic acid.

First claim

The invention claimed is:

1. A method for enhancing immunogenicity of a target protein, comprising fusing the target protein to a fragment of CRM197, optionally via a linker, to obtain a fusion protein, wherein the fragment of CRM197 consists of aa 1-190 or aa 1-389 of SEQ ID NO: 2, and wherein the fragment of CRM197 enhances immunogenicity of the target protein.

2. The method of claim 1 , wherein the fusion protein is obtained by fusion expression of the fragment of CRM197 with the target protein, optionally using a linker.

3. The method of claim 1 , wherein the fusion protein comprises the fragment of CRM197 which is linked to the N-terminus and/or C-terminus of the target protein, optionally via a linker.

4. The method of claim 1 , wherein the target protein is HEV capsid protein or an immunogenic fragment thereof.

5. The method of claim 4 , wherein the immunogenic fragment of HEV capsid protein comprises or is HEV-239 (aa 368-606 of HEV capsid protein), E2 (aa 394-606 of HEV capsid protein) or E2s (aa 455-606 of HEV capsid protein).

6. The method of claim 4 , wherein the fusion protein comprises (a) the fragment of CRM197, and (b) HEV capsid protein or the immunogenic fragment of HEV capsid protein; wherein (a) and (b) are linked together, optionally via a linker.

7. The method of claim 4 , wherein the fusion protein has an amino acid sequence as set forth in SEQ ID NO: 8, 10, 12, 14, 16 or 18.

8. The method of claim 1 , wherein the target protein is influenza virus M2 protein or an immunogenic fragment thereof.

9. The method of claim 8 , wherein the immunogenic fragment of M2 protein comprises or is M2e (aa 1-24 of M2 protein).

10. The method of claim 8 , wherein the fusion protein comprises (a) the fragment of CRM197, and (b) influenza virus M2 protein or the immunogenic fragment of M2 protein; wherein (a) and (b) are linked together, optionally via a linker.

11. The method of claim 8 , wherein the fusion protein has an amino acid sequence as set forth in SEQ ID NO: 36, 38, 42 or 44.

12. The method of claim 1 , wherein the fusion protein comprises a linker.

Assignees

• Univ Xiamen
• Xiamen Innovax Biotech Co Ltd

Inventors

• Li Shaowei
• Song Cuiling
• Yang Chunyan
• Gu Ying
• Luo Wenxin
• Xia Ningshao

Classifications

• A61P37/04

  Immunostimulants · CPC title

• A61P31/16

  for influenza or rhinoviruses · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P31/14

  for RNA viruses · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title