Rep protein for use in a diagnostic assay

Patent metadata

Field	Value
Publication number	US-10907141-B2
Application number	US-201916730473-A
Country	US
Kind code	B2
Filing date	Dec 30, 2019
Priority date	Jul 7, 2017
Publication date	Feb 2, 2021
Grant date	Feb 2, 2021

Abstract

The present invention concerns a DNA-replication-associated (Rep) protein comprising an amino acid sequence as depicted in SEQ ID NO: 11 or 12; (b) a fragment of SEQ ID NOs:11 or 12 which is capable of binding an anti-Rep antibody specific for a protein having the amino acid sequence of SEQ ID NOs: 11 or 12; or (c) an amino acid sequence having a 90% or more homology to the amino acid sequence of (a) or (b) and is capable of binding an anti-Rep antibody specific for a protein having an amino acid sequence of SEQ ID NOs:11 or 12. The present invention further concerns a method of diagnosing MS or a predisposition for MS and a kit for use in such methods.

First claim

The invention claimed is:

1. A DNA-replication-associated (Rep) protein comprising the amino acid sequence of SEQ ID NOs: 11 or 12.

2. A method of diagnosing multiple sclerosis (MS) in a subject comprising the steps of
(a) incubating a sample from a subject with a Rep protein as defined in claim 1 ;
(b) detecting the amount of antibodies in the sample from the subject forming an immunological complex with the Rep protein; and
(c) correlating the amount of antibody bound to Rep protein in the sample from the subject, as compared to an amount in a control sample, for diagnosing MS.

3. The method of claim 2 , wherein in step (a) the Rep protein is immobilized followed by incubating the immobilized Rep protein with the sample from the subject.

4. The method of claim 2 , wherein in step (a) the Rep protein is expressed in cells followed by incubating the cells with the sample from the subject.

5. The method of claim 2 , wherein in step (b) the amount of antibodies forming an immunological complex with Rep protein is quantified by a detecting binding agent coupled to a signal generating compound.

6. The method of claim 2 , wherein in step (a), the antibodies in the sample from the subject are immobilized prior to incubating with a defined amount of the Rep protein.

7. The method of claim 2 , wherein the sample from the subject is a serum or a plasma sample.

8. The method of claim 2 , wherein MS or a predisposition for MS is indicated by an increased amount of anti-Rep antibodies in the subject's sample of at least 2 fold as compared to a control sample.

9. A kit for use in the diagnosis of MS comprising:
(a) a Rep protein, wherein the Rep protein comprises the amino acid sequence of SEQ ID NO:11 or 12;
(b) an anti-human antibody coupled to a detectable label and capable of binding to anti-Rep antibody with a specificity for a protein having the amino acid sequence of SEQ ID NO: 11 or 12, and
(c) a solid matrix suitable for immobilizing a Rep protein according to (a).

10. The kit according to claim 9 for use in an assay selected from the group consisting of enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), enzyme immune assay (EIA), fluorescence immunoassay (FIA), luminescence immunoassay (LIA) and strip assay.

Assignees

• Deutsches Krebsforsch

Inventors

• Bund Timo
• De Villiers-Zur Hausen Ethel-Michele
• Zur Hausen Harald

Classifications

• G01N2800/285

  Demyelinating diseases; Multipel sclerosis · CPC title

• G01N33/564

  for pre-existing immune complex or autoimmune disease {, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9} · CPC title

• C07K14/4713 Primary

  Autoimmune diseases, e.g. Insulin-dependent diabetes mellitus, multiple sclerosis, rheumathoid arthritis, systemic lupus erythematosus; Autoantigens · CPC title

• C07K16/18

  against material from animals or humans · CPC title

• G01N33/577

  involving monoclonal antibodies {binding reaction mechanisms characterised by the use of monoclonal antibodies (G01N33/5302 - G01N33/576 take precedence)} · CPC title