Complexes comprising a prion protein and a peptidyl prolyl isomerase chaperone, and method for producing and using them
AssigneeRoche Diagnostics GmbH
InventorsElke Faatz, Christian Scholz, Werner Stock, Peter Schaarschmidt
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Abstract
The present invention relates to the diagnosis of transmissible spongiform encephalopathies (TSEs). It especially teaches the production of a soluble prion protein (PrP)-chaperone complex and the advantageous use of such chaperone-PrP complex, especially in the detection of PrP in an immunoassay, as well as its use as and immunogen.
Claims65
A protein which is soluble to at least 100 nM in a solution which has a pH of 7.4 and consists of 20 nM sodium phosphate and 150 nM sodium chloride, said protein comprising:
- a prion protein (PrP), and
- a peptidyl prolyl isomerase chaperone,
- wherein the PrP and the peptidyl prolyl isomerase chaperone are covalently linked.
A recombinant polypeptide which is soluble to at least 100 nM in a solution which has a pH of 7.4 and consists of 20 nM sodium phosphate and 150 nM sodium chloride, said recombinant polypeptide comprising:
- a prion protein (PrP),
- a peptidic linker, and
- a peptidyl prolyl isomerase chaperone.
A recombinantly-produced fusion polypeptide comprising:
- a prion protein (PrP), and
- an FKBP chaperone polypeptide, wherein said FKBP chaperone polypeptide is selected from the group consisting of FkpA, SlyD, and trigger factor.
A recombinant polypeptide which is soluble to at least 100 nM in a solution which has a pH of 7.4 and consists of 20 nM sodium phosphate and 150 nM sodium chloride, said recombinant polypeptide comprising:
- at least one PrP domain of a prion protein (PrP),
- a peptidic linker, and
- at least one chaperone domain of a peptidyl prolyl isomerase chaperone.
An expression vector comprising:
- at least one nucleic acid sequence encoding a prion protein (PrP),
- at least one nucleic acid sequence encoding a peptidic linker, and
- at least one nucleic acid sequence encoding an FKBP chaperone selected from the group consisting of FkpA, SlyD, and trigger factor.
An expression vector comprising:
- at least one nucleic acid sequence encoding a prion protein (PrP),
- at least one nucleic acid sequence encoding a peptidic linker, and
- at least one nucleic acid sequence encoding a peptidyl prolyl isomerase chaperone.
An expression vector comprising:
- at least one nucleic acid sequence encoding at least one PrP domain of a prion protein (PrP),
- at least one nucleic acid sequence encoding a peptidic linker, and
- at least one nucleic acid sequence encoding at least one chaperone domain of a peptidyl prolyl isomerase chaperone.
A method for producing a soluble prion protein (PrP)-chaperone protein comprising:
- incubating a polypeptide comprising PrP covalently linked to a peptidyl prolyl isomerase chaperone in a buffer wherein both the PrP and the chaperone are solubilized to at least 100 nM, said buffer comprising a pH of 7.4, 20 mM sodium phosphate, and 150 mM sodium chloride.
Dependencies are inferred from the claim text.