Polypeptides with affinity for heat shock proteins (HSPS) and HSP associated complexes (HACS) and their use in diagnosis and therapy
The present application is directed to a peptides comprising an a-helix forming-amino acid sequence that binds a heat shock protein. Also included is a polypeptide comprising (a) a first peptide portion that comprises an α-helix-forming amino acid sequence that binds a heat shock protein; and (b) at least one second peptide portion comprising an antigenic amino acid sequence and/or an a-helix-stabilizing amino acid sequence that increases the interaction of the first peptide portion with the heat shock protein. The present application also includes compositions comprising the peptides and/or polypeptides of present application and uses of the peptides and/or polypeptides of the present application for fractionating substances relevant for discovery, research or clinical analysis from a biological sample and as therapeutics.
1. A polypeptide comprising:
(a) a first peptide portion consisting of an α-helix-forming amino acid sequence of myelin basic protein having the amino acid sequence of PSQGKGRGLSLSRFSWGA [SEQ ID NO 2:] or a sequence having one amino acid deletion, addition or substitution relative to [SEQ ID NO 2]; and
(b) at least one second peptide portion comprising a tumour associated antigen (TAA) epitope of survivin at least 8 amino acids in length or a sequence having one amino acid deletion, or substitution relative to the epitope of survivin.
2. The polypeptide of claim 1 , wherein the at least one second peptide portion comprises one or more of:
[SEQ ID NO: 7]
STFKNWPFL,
[SEQ ID NO: 8]
LTLGEFLKL;
or
[SEQ ID NO: 9]
LRRKCAVPS
or a sequence having one amino acid deletion,
addition or substitution relative to [SEQ ID NO:
7], [SEQ ID NO: 8] or [SEQ ID NO: 9].
3. The polypeptide of claim 1 , wherein the first peptide portion is at the N-terminus of the polypeptide and comprises one second peptide portion at the C-terminus.
4. The polypeptide of claim 1 , wherein the first peptide portion of the polypeptide is located between two second peptide portions, which is the same or different.
5. The polypeptide of claim 1 , comprising an amino acid sequence selected from:
[SEQ ID NO: 64]
PSQGKGRGLSLSRFSWGASTFKNWPFL;
[SEQ ID NO: 65]
PSQGKGRGLSLSRFSWGALTLGEFLKL;
and
[SEQ ID NO: 66]
PSQGKGRGLSLSRFSWGALRRKCAVPS
or a sequence having one or two amino acid
deletions, additions or substitutions relative to
[SEQ ID NO: 64], [SEQ ID NO: 65] or [SEQ ID NO:
66]
6. The polypeptide of claim 1 , further comprising a linker moiety, a secondary modification or label.
7. A nucleic acid encoding the polypeptide of claim 1 .
8. A composition comprising the polypeptide of claim 1 , and one or more additives, excipients and/or adjuvants.
9. A method of fractionating one or more substances relevant for discovery, research or clinical analysis from a biological sample comprising: (a) contacting the biological sample with one or more of the polypeptides of claim 1 under conditions suitable for binding of the one or more polypeptides to the one or more substances to form complexes, and (b) fractionating the complexes.
10. A method of identifying substances for diagnosing an infectious disease or cancer in a subject comprising: (a) contacting a sample from the subject with one or more polypeptides of claim 1 under conditions suitable for the binding of the of the one or more peptides or polypeptides to one or more infectious disease and/or cancer relevant substances to form complexes, (b) fractionating the complexes, and (c) identifying the presence of the one or more infectious disease and/or cancer relevant substances in the complexes, wherein the one or more infectious disease and/or cancer relevant substances are comprised in an HSP associated complex.
11. A method of inducing an immune response comprising administering an effective amount of one or more of the polypeptide of claim 1 to a subject in need thereof.
12. The polypeptide of claim 1 , wherein the first peptide portion comprises the sequence: PSQGKGRGLSLSRFSWGA [SEQ ID NO 2:].
13. The polypeptide of claim 2 , wherein the TAA comprises the amino acid sequence of: LTLGEFLKL [SEQ ID NO:8].
14. The polypeptide of claim 1 , comprising an amino acid sequence at least 90% identical to PSQGKGRGLSLSRFSWGALTLGEFLKL [SEQ ID NO: 65].
15. The polypeptide of claim 1 , further comprising one of more of a phosphorylation, acylation, acetylation, formylation, glycosylation, amidation, incorporation of chromophores, fluorescent dyes/fluorogenic groups, PEGylation, biotinylation and sulfation.
16. The polypeptide of claim 1 , wherein the polypeptide is linked to a solid support.
17. A complex comprising the polypeptide of claim 1 and a heat shock protein, wherein the polypeptide and heat shock protein are affinity associated.
18. The complex of claim 17 wherein the heat shock protein is selected from HSP60, HSP70, HSP90 or HSP27 or an isoform thereof.
19. A vector comprising the nucleic acid of claim 7 .
20. The composition of claim 8 , wherein the one or more additives comprises a detergent matrix.
21. The method of claim 9 , wherein the one or more substances relevant for discovery, research or clinical analysis are proteins.
22. The method of claim 9 , wherein the one or more substances relevant for discovery, research or clinical analysis are cell derived vesicles (CDVs).
23. The method of claim 9 , wherein the one or more substances relevant for discovery, research or clinical analysis are HSPs or HSP-associated complexes.
24. The method of claim 23 , wherein the HSP is HSP60, HSP70, HSP90 or HSP27 or an isoform thereof.
25. The method of claim 9 , wherein the biological sample is a bodily fluid.
26. The method of claim 25 , wherein the bodily fluid is selected from blood, plasma, urine, cerebrospinal fluid, lymph, ascites, saliva, lavage, semen, glandular secretions, feces, exudates and contents of cysts.
27. The method of claim 9 , wherein the conditions suitable for binding of the one or more polypeptides to the one or more substances to form complexes comprise counter affinity isoelectric focusing.
28. The method of claim 9 , wherein the conditions suitable for binding of the one or more polypeptides to the one or more substances to form complexes comprise contacting solutions of the one or more peptides with the biological sample in a biologically relevant solution and optionally applying agitation, heat and/or microwaves.
29. The method of claim 9 , wherein the one or more substances relevant for discovery, research or clinical analysis are substances for diagnosing an infectious disease or cancer in a subject.
30. A kit comprising the one or more of the polypeptides claim 1 and a dilution buffer.