Rep protein as protein antigen for use in diagnostic assays

Patent metadata

Field	Value
Publication number	US-11718662-B2
Application number	US-201916376154-A
Country	US
Kind code	B2
Filing date	Apr 5, 2019
Priority date	Oct 10, 2016
Publication date	Aug 8, 2023
Grant date	Aug 8, 2023

Abstract

Disclosed is a method of diagnosing multiple sclerosis (MS), wherein a blood sample from a patient is incubated with a DNA-replication associated (REP) protein. The present invention relates to a DNA-replication-associated (Rep) protein for use in the diagnosis of multiple sclerosis (MS), wherein (a) an increased amount of Rep protein or fragments thereof in the sample as compared to an amount in a control sample; or an increased amount of anti-Rep protein antibodies with antigen in a sample from a subject as compared to an amount in a control sample correlates with a diagnosis of MS, wherein the Rep protein is MSBI1 Rep or MSBI2 Rep.

First claim

What is claimed is:

1. A method of diagnosing multiple sclerosis (MS) in a human patient comprising the steps of
(a) incubating a sample from the human patient selected from the group consisting of blood, serum and plasma with a DNA-replication-associated (Rep) protein comprising,
(i) the amino acid sequence as depicted in SEQ ID NOS: 1 or 14; or
(ii) a fragment of the amino acid sequence as depicted in SEQ ID NO: 1 and the fragment consists of the amino acid selected from the group consisting of the amino acids 1 to 136 or 137 to 229 of SEQ ID NO:1 and SEQ ID NOS: 2, 3 or 9;
(b) detecting the amount of antibodies in the sample from the human patient forming an immunological complex with the Rep protein;
(c) determining the extent of difference in the amount of antibodies present in the sample from the human patient and a control sample taken from a healthy individual who has not been diagnosed for MS; and
(d) diagnosing MS, when the amount of antibodies bound to Rep protein in the sample from the human patient is at least 2-fold increased, as compared to the amount in the control sample.

2. The method of claim 1 , wherein the protein is encoded by a polynucleotide acid as depicted in SEQ ID NO:13.

3. The method of claim 1 , 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 1 , 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 3 , wherein in step (b) the amount of antibodies forming an immunological complex with the immobilized Rep protein is quantified by a detecting binding agent coupled to a signal generating compound.

6. The method of claim 4 , 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.

7. The method of claim 1 , wherein in step (a) the antibodies in the sample from the subject are immobilized followed by incubating with a defined amount of Rep protein.

8. The method of claim 5 , wherein in step (a) the Rep protein is bound to a solid support and in step (b) the detecting binding agent is an anti-human antibody coupled to a signal generating compound.

9. The method of claim 8 , wherein the signal generating compound is selected from the group consisting of enzymatic, fluorescent, chemiluminescent, radioactive and dye molecules.

10. The method of claim 9 , wherein the signal generating compound is horseradish peroxidase.

11. The method of claim 4 , wherein the Rep protein expressed in cells comprises the amino acid sequence as depicted in SEQ ID NO: 14.

12. The method of claim 11 , wherein the Rep protein is encoded by the polynucleotide acid as depicted in SEQ ID NO: 13.

Assignees

• Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts
• Deutsches Krebsforschungszentrum Stiftung Des Oeffentlichen Rechts

Inventors

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

Classifications

• C07K16/18 Primary

  against material from animals or humans · CPC title

• G01N33/502

  for testing non-proliferative effects · CPC title

• G01N33/53 Primary

  Immunoassay; Biospecific binding assay; Materials therefor · CPC title

• G01N33/563

  involving antibody fragments · CPC title

• G01N33/564 Primary

  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