Use of prion conversion modulating agents
View Patent ↗The use of Apolipoprotein B, Apolipoprotein E, fragments and mimetics thereof is provided for diagnostic, detection, prognostic and therapeutic applications In prion diseases. More specifically, the invention provides the use of Apolipoprotein B or fragments thereof for modulating or identifying modulators of the prion protein replication which are implicated in the pathogenesis of transmissible spongiform encephalopathics and other prion diseases.
1. A method for the detection of a prion disease within a subject suspected of suffering from such a disease, the method comprising:
(i) contacting a sample from said subject with Apolipoprotein B or a peptide fragment thereof;
(ii) contacting the preparation obtained in step (i) with PrP C or a PrP C containing mixture; and
(iii) determining the presence and/or an amount of PrP Sc in said sample;
wherein the presence of PrP Sc in said sample is indicative of the presence of prions in said subject.
2. The method of claim 1 , wherein the prion disease is bovine spongiform encephalopathy (BSE).
3. The method of claim 1 , wherein the prion disease is a Creutzfeld-Jacob disease.
4. A method for the detection of PrP Sc within a sample, comprising:
(i) contacting said sample with Apolipoprotein B or a peptide fragment thereof;
(ii) contacting the sample obtained in (i) with PrP C or a PrP C containing mixture; and
(iii) determining the presence and/or an amount of PrP Sc in said sample,
wherein the presence of PrP Sc indicates that the sample contains PrP Sc .
5. A method for identifying, in a sample, a compound which modulates the transition of PrP C into PrP Sc , comprising:
(i) contacting said sample with Apolipoprotein B or a peptide fragment thereof; in the presence of said modulatory compound and (b) in the absence of said compound;
(ii) contacting the preparation obtained in step (i) a and (i) b with PrP C or a PrP C containing mixture; and
(iii) determining the amount of PrP Sc (a) in the presence of said modulatory compound and (b) in the absence of said modulatory compound,
wherein the presence of PrP Sc identifies a compound that modulates the transition of PrP C into PrP Sc .
6. The method of claim 1 , wherein the peptide or the protein contains the sequence of SEQ ID NO: 3.
7. The method of claim 1 , wherein the peptide or the protein has a molecular weight from 30 and 40 kDa and has a sequence selected from the group of Apolipoprotein B between positions 3201-3558, 3548-3905, 3201-3905, 3291-3558, 3548-3815, and 3291-3815.
8. The method of claim 1 , wherein the protein is Apolipoprotein B or a fragment thereof consisting of a peptide fragment of Apolipoprotein B between positions 3201-3558, 3548-3905, 3201-3905, 3291-3558, 3548-3815, or 3291-3815.
9. The method of claim 1 , wherein the peptide or the protein forms a complex with a LDL receptor.
10. The method of claim 1 , wherein the peptide or the protein contains the sequence of SEQ ID NO: 3.
11. The method of claim 1 , wherein the peptide or the protein has a molecular weight from 30 and 40 kDa and is a fragment of Apolipoprotein B comprising the consecutive amino acid residues between positions 3201-3553, 3548-3905, 3201-3905, 3291-3558, 3548-3815, or 3291-3815.
12. The method of claim 4 , wherein the protein is Apolipoprotein B or a fragment thereof consisting of a peptide fragment of Apolipoprotein B between positions 3201-3558, 3548-3905, 3201-3905, 3291-3558, 3548-3815, or 3291-3815.
13. The method of claim 4 , wherein the peptide or the protein from a complex wit a LDL receptor.
14. The method of claim 4 , wherein the peptide or the protein contains the sequence of SEQ ID NO: 3.
15. The method of claim 4 , wherein the peptide or the protein has a molecular weight from 30 and 40 kDa and is a fragment of Apolipoprotein B comprising the consecutive amino acid residues between positions 3201-3558, 3548-3905, 3201-3905, 3291-3558, 3548-3815, or 3291-3815.
16. The method of claim 4 , wherein the sample is obtained from a subject suspected of having a prion disease selected from the group consisting of bovine spongiform encephalopathy (BSE) and Creutzfeld-Jacob Disease (CJD).
17. The method of claim 5 , wherein the protein is Apolipoprotein B or a fragment thereof consisting of a peptide fragment of Apolipoprotein B between positions 3201-3558, 3548-3905, 3201-3905, 3291-3558, 3548-3815, or 3291-3815.
18. The method of claim 5 , wherein the peptide or the protein forms a complex with a LDL receptor.
19. The method of claim 5 , wherein the peptide or the protein contains the sequence of SEQ ID NO: 3.
20. The method of claim 5 , wherein the peptide or the protein has a molecular weight from 30 and 40 kDa and is a fragment of Apolipoprotein B comprising the consecutive amino acid residues between positions 3201-3558, 3548-3905, 3201-3905, 3291-3558, 3548-3815, or 3291-3815.
21. The method of claim 5 , wherein the sample is obtained from a subject suspected of having a prion disease selected from the group consisting of bovine spongiform encephalopathy (BSE) and Creutzfeld-Jacob Disease (CJD).
22. The method of claim 5 , wherein determining the amount of PrP Sc in the sample comprises performing a protein misfolding cyclic amplification (PMCA) assay.
23. The method of claim 22 , wherein the sample is a normal brain homogenate containing PrP C and substrate.
24. The method of claim 22 , wherein the sample is lipid rafts from an infection-sensitive neuroblasma cell line N2a containing PrP C and substrate.
25. The method of claim 5 , which comprises determining the amount of PrP Sc in the sample by performing a protein misfolding cyclic amplification assay (PMCA); and
wherein the protein is Apolipoprotein B, and
wherein the sample is lipid rafts from infection sensitive neuroblasma cell line N2a that contain normal PrP C and substrate.
26. The method of claim 5 , wherein said modulatory compound is an antagonist of Apolipoprotein B.
27. The method of claim 5 , wherein said modulatory compound is an antibody that binds to Apolipoprotein B.
28. The method of claim 5 , wherein said modulatory compound is a LDL-receptor antagonist.
29. A method for the detection of PrP Sc in a biological sample comprising:
(i) contacting a sample from a subject exposed to a prion with Apolipoprotein B or a peptide fragment thereof;
(ii) contacting the preparation obtained in step (i) with PrP C or PrP C containing mixtures; and
(iii) determining the presence and/or an amount of PrP Sc in said sample,
wherein the presence of PrP Sc in said sample is indicative of the presence of prions in the sample.
30. The method of claim 29 , wherein said subject is human and has, or is at risk of developing, vCJD.