IgG binding peptides and multifunctional antigen-binding proteins comprising the same

Patent metadata

Field	Value
Publication number	US-11312784-B2
Application number	US-201716309969-A
Country	US
Kind code	B2
Filing date	Jun 16, 2017
Priority date	Jun 17, 2016
Publication date	Apr 26, 2022
Grant date	Apr 26, 2022

Abstract

Disclosed herein are multifunctional antigen-binding proteins comprising at least one multifunctional recombinant protein scaffold and at least one antigen-specific binding domain. Polynucleotides encoding the multifunctional antigen-binding proteins, vectors containing the disclosed polynucleotides, and cells that have been genetically engineered to express the polynucleotide are also provided. Methods of using the multifunctional antigen-binding proteins are disclosed.

First claim

What is claimed:

1. A multifunctional antigen-binding protein comprising, at least one multifunctional recombinant protein scaffold that binds to IgG and at least one antigen-specific binding domain,
wherein the at least one multifunctional recombinant protein scaffold that binds to IgG comprises a human polypeptide that contains mutations that enable the polypeptide to mimic the IgG binding domain of protein A,
wherein the at least one multifunctional recombinant protein scaffold comprises a polypeptide that is:
(a) at east 90% identical to huZZ as set forth in SEQ ID NO:28; or
(b) at least 90% identical to huZZ1 as set forth in SEQ ID NO:29; and
wherein the at least one antigen-specific binding domain comprises an antigen-specific peptide, an antigen-specific antibody or fragment thereof, or a combination thereof.

2. The multifunctional antigen-binding protein of claim 1 , wherein the at least one multifunctional recombinant protein scaffold that binds to IgG comprises a portion of an alpha2-macroglobulin receptor-associated protein.

3. The multifunctional antigen-binding protein of claim 1 , wherein the at least one multifunctional recombinant protein scaffold has the sequence set forth as SEQ ID NO:28 or SEQ ID NO:29.

4. The multifunctional antigen-binding protein of claim 1 , wherein the at least one multifunctional recombinant protein scaffold is linked to said antigen-specific binding domain and is capable of binding to a protein other than an antigen which can be hound by the antigen-specific binding domain.

5. The multifunctional antigen-binding protein of claim 1 , wherein the antigen-specific peptide comprises a polypeptide that is at least 90% identical to any one of SEQ ID NOs:12-24.

6. The multifunctional antigen-binding protein of claim 1 , wherein the antigen-specific antibody comprises a polypeptide that is at least 90% identical to 4D5scFv as set forth in SEQ ID NO:30.

7. The multifunctional antigen-binding protein of claim 6 , said multifunctional antigen-binding protein having an amino acid sequence that is:
(a) at least 90% identical to 4D5scFv-huZZ as set forth in SEQ NO:31; or
(b) at least 90% identical to 4D5scFv-huZZ1 as set forth in SEQ NO:32.

8. The multifunctional antigen-binding protein of claim 1 , further comprising a protein domain that promotes solubility.

9. The multifunctional antigen-binding protein of claim 1 , further comprising an epitope tag, a fluorophore, a radio isotope, an enzyme, or any combination thereof.

10. The multifunctional antigen-binding protein of claim 9 , wherein said epitope tag is a poly-histidine tag.

11. The multifunctional antigen-binding protein of claim 1 , further comprising a linker having one or more glycine residues, wherein said linker connects the at least one multifunctional recombinant protein scaffold to the at least one antigen-specific binding domain.

12. The multifunctional antigen-binding protein of claim 11 , wherein the linker comprises a polypeptide that is at least 90% identical to SEQ ID NO:25 or SEQ ID NO:26.

13. The multifunctional antigen-binding protein of claim 1 , further comprising an interferon (IFN) polypeptide or portion thereof.

14. The multifunctional antigen-binding protein of claim 13 , said multifunctional antigen-binding protein having an amino acid sequence that is:
(a) at least 90% identical to 4D5scFv-huZZ-hIFN as set forth in SEQ ID NO:33;
(b) at least 90% identical to 4D5scFv-huZZ-mIFN as set forth in SEQ ID NO:34;
(c) at least 90% identical to 4D5scFv-huZZ1-hIFN as set forth in SEQ ID NO:35; or
(d) at least 90% identical to 4D5scFv-huZZ1-mIFN as set forth in SEQ NO:36.

15. A polynucleotide encoding the multifunctional antigen-binding protein of claim 1 .

16. A cell that has been genetically engineered to express the polynucleotide of claim 15 .

17. A composition comprising the multifunctional antigen-binding protein of claim 1 and a pharmaceutically acceptable carrier.

18. A polypeptide comprising the sequence as set forth in SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ NO:34, SEQ ID NO:35, or SEQ ID NO:36.

19. A polynucleotide encoding the polypeptide of claim 18 .

Assignees

• Univ Pennsylvania

Inventors

• Greene Mark I
• Zhang Hongtao
• Zhu Zhiqiang
• Lam Lian
• Cai Zheng

Classifications

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title

• C07K14/195

  from bacteria · 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

• C07K2319/21

  containing a His-tag · CPC title