Methods and compositions involving fibrillizing polypeptides for nanofibers

• US 9,200,082 B2
• Filed: 03/13/2014
• Issued: 12/01/2015
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: University of Chicago
• Original Assignee: University of Chicago
• Inventors: Collier, Joel, Hudalla, Gregory
• Primary Examiner(s): Skowronek, Karlheinz R
• Assistant Examiner(s): NEWMAN, MINDY B
• Application Number:
  US14/209,757
• Publication Number:
  US 20140273148A1
• Time in Patent Office: 628 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 17/06 attached to the carrier via a bridging agent

Abstract

Embodiments of the invention are directed to fibrillar adjuvants. For example, epitopes assembled by a synthetic peptide domain into nanofibers comprising a β-fibrillization peptide may elicit high antibody titers in the absence of any adjuvant. In certain embodiments, multiple different antigens may be integrated into polypeptide nanofibers, providing biomaterials with modular and precise composition of bioactive proteins.

Claims

1. A nanofiber complex composition, wherein the composition comprises a β

-sheet nanofiber structure comprising

a) a plurality of non-β

-sheet peptide tags that undergo a transition from a non-β

-sheet structure to a β

-sheet structure in the presence of β

-sheet peptides, wherein a non-β

-sheet peptide tag is attached to a compound; and

b) a plurality of β

-sheet peptides.

2. The composition of claim 1, wherein the structure comprises at least two different compounds.

3. The composition of claim 1, wherein the non-β

-sheet peptides tags are α

-helical peptides.

4. The composition of claim 1, wherein non-β

-sheet peptides tags comprise one or more alpha helical motifs having a sequence of a b c de f g, with a and d being non-polar amino acids and e and g being charged amino acids.

5. The composition of claim 4, wherein a and/or d is Ala (A), Leu (L), Ile (I), Val (V) or a conservative derivative thereof in one or more of the alpha helical motifs.

6. The composition of claim 4, wherein a and/or d is Leu (L) in one or more of the alpha helical motifs.

7. The composition of claim 4, wherein e and/or g is Lys (K), Arg (R), His (H), Asp (D), Glu (E) or a conservative derivative thereof in one or more of the alpha helical motifs.

8. The composition of claim 4, wherein one or more of b, c, and f is a hydrophobic amino acid in one or more of the alpha helical motifs.

9. The composition of claim 4, wherein one or more of b, c, and f in one or more of the alpha helical motifs is Val (V), Tyr (Y), Phe (F), Trp (W), Ile (I), or Thr (T).

10. The composition of claim 4, wherein one or more of b, c, and f is Val (V) in one or more of alpha helical motifs.

11. The composition of claim 1, wherein the non-β

-sheet peptide tag comprises an amino acid sequence having at least 90% identity with the sequence of LVVLHSELHKLKSEL (SEQ ID NO;

     1), LVVLHSHLEKLKSEL (SEQ ID NO;

     2), LKVELEKLKSELVVLHSELHKLKSEL (SEQ ID NO;

     3), LKVELEKLKSELVVLHSHLEKLKSEL (SEQ ID NO;

     4), or LKVELKELKKELVVLKSELKELKKEL (SEQ ID NO;

     5).

12. The composition of claim 1, wherein one or more of the alpha helical motifs further comprise at least two metal binding amino acids spaced by one or three amino acids.

13. The composition of claim 1, wherein the non-β

-sheet peptide tag has 14 to 56 amino acids in length.

14. The composition of claim 1, wherein the compound attached to the non-β

-sheet peptide tags is a peptide, polypeptide, nucleic acid, small molecule, antigen, ligand, enzyme, reporter, drug, matrix, cell, virus, bacterium, lipid, carbohydrate, or a combination thereof.

15. The composition of claim 1, wherein at least one of the non-β

-sheet peptide tags attached to a compound is a fusion protein.

16. The composition of claim 15, wherein one or more of the non-β

-sheet peptide tags are attached to the amino-terminus of a peptide.

17. The composition of claim 1, wherein the compound attached to a non-β

-sheet peptide tag is an enzyme, fluorescent protein, cell binding domain, cell adhesion domain, extracellular matrix domain, reporter protein, cytokine, antigen, signaling domain, immunomodulating protein, cross-linking protein, hormone, hapten, or a combination thereof.

18. The composition of claim 1, wherein the β

-sheet peptides comprise a plurality of self-assembling peptides.

19. The composition of claim 1, wherein the β

-sheet peptide has 2 to 40 amino acids in length.

20. A method of preparing a nanofiber complex composition of claim 1, comprising mixing the following:

a) a plurality of non-β

-sheet peptide tags, wherein a non-β

-sheet peptide tag is attached to a compound; and

b) a plurality of β

-sheet peptides, under conditions that allow one or more of the non-β

-sheet peptide tags to undergo transition from a non-β

-sheet structure to a β

-sheet structure, thereby preparing a nanofiber complex composition that forms a β

-sheet structure comprising the transitioned non-β

-sheet peptide tags and β

-sheet peptides.