Modified biotin-binding protein, fusion proteins thereof and applications

Patent metadata

Field	Value
Publication number	US-9499593-B2
Application number	US-201214116492-A
Country	US
Kind code	B2
Filing date	May 11, 2012
Priority date	May 11, 2011
Publication date	Nov 22, 2016
Grant date	Nov 22, 2016

Abstract

The disclosure provides modified biotin-binding proteins which can be expressed in soluble form in high yield in bacteria. Also provided are fusion proteins comprising the modified biotin-binding protein and an antigen. The disclosure further provides non-hemolytic variants of alpha-hemolysin from S. aureus and fusion protein comprising non-hemolytic variant of alpha-hemolysin and a biotin-binding domains. Immunogenic compositions comprising the proteins are also disclosed and use of such immunogenic compositions for inducing an immune response or for vaccinating a subject are also disclosed.

First claim

We claim:

1. A vaccine composition comprising a soluble biotin-binding protein consisting essentially of an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1, wherein the biotin-binding protein has a higher solubility than the biotin-binding protein of SEQ ID NO: 4.

2. The vaccine composition of claim 1 , wherein the soluble biotin-binding protein further comprises one or more of: a signal sequence, a lipidation sequence, a purification tag, a linker peptide, and a spacer peptide.

3. An immunogenic composition comprising at least one soluble biotin-binding protein, wherein the soluble biotin-binding protein consists essentially of an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1, wherein the biotin-binding protein has a higher solubility than the biotin binding protein of SEQ ID NO: 4, and wherein the soluble biotin-binding protein optionally further comprises one or more of: a signal sequence, a lipidation sequence, a purification tag, a linker peptide, and a spacer peptide.

4. The immunogenic composition of claim 3 , further comprising at least one or more of: a polysaccharide, and a polypeptide.

5. The immunogenic composition of claim 4 , comprising at least one polysaccharide and at least one polypeptide.

6. The immunogenic composition of claim 5 , wherein the at least one polysaccharide is associated with a first affinity molecule.

7. The immunogenic composition of claim 5 , wherein the soluble biotin-binding protein is non-covalently associated with the at least one polypeptide.

8. The immunogenic composition of claim 5 , wherein the at least one polysaccharide is associated with a first affinity molecule, and the soluble biotin-binding protein is non-covalently associated with the at least one polypeptide.

9. The immunogenic composition of claim 8 , wherein the first affinity molecule associates with the soluble biotin-binding protein to link the polypeptide and the polysaccharide.

Assignees

• Malley Richard
• Lu Yingjie
• Zhang Fan
• Children'S Medical Center Corp

Inventors

• Malley Richard
• Lu Yingjie
• Zhang Fan

Classifications

• A61K2039/627

  characterised by the linker · CPC title

• A61K2039/6093

  Synthetic polymers, e.g. polyethyleneglycol [PEG], Polymers or copolymers of (D) glutamate and (D) lysine · CPC title

• A61K2039/55505

  Inorganic adjuvants · CPC title

• A61K2039/60

  characteristics by the carrier linked to the antigen · CPC title

• C07K14/31 Primary

  from Staphylococcus (G) · CPC title