# US 2014/0154709 A1 · Methods of measuring adamts 13-mediated in vivo cleavage of von willebrand factor and uses thereof

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2014/0154709 A1
- Document ID: US20140154709A1
- Status: Published application
- Kind: A1
- Application number: 14/078,324
- Filed: 2013-11-12
- Published: 2014-06-05
- Earliest claimed: 2008-12-05
- Assignees: Baxter Healthcare S.A.; Baxter Internationsl Inc.
- Applicants: Baxter Healthcare SA; Baxter Internationsl Inc.
- Inventors: Katalin Varadi; Hanspeter Rottensteiner; Peter Turecek; Hans-Peter Schwarz; Jutta Schreiner
- Web page: https://patentgrep.com/patent/US20140154709A1

## CPC

- G01N 33/86: involving blood coagulating time {or factors, or their receptors}
- C12Q 1/37: involving peptidase or proteinase
- C12Q 1/56: involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
- G01N 33/573: for enzymes or isoenzymes
- G01N 2333/755: Factors VIII, e.g. factor VIII C [AHF], factor VIII Ag [VWF]
- G01N 2333/96486: Metalloendopeptidases (3.4.24)
- G01N 2800/224: Haemostasis or coagulation
- G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The invention generally relates to methods of measuring cleaved von Willebrand factor (VWF) fragments. More specifically, the invention relates to methods of measuring the ability of a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) to cleave VWF in vivo. The invention also relates to methods of using various animal models which demonstrate ADAMTS13 activity similar to that of a human. The invention further relates to methods of measuring the cleavage products of rVWF in mammals, particularly in humans and in human plasma.

## Claims

### Claim at position 1

1-6. (canceled)

### Claim 7

A method for testing the effectiveness of a treatment for thrombotic thrombocytopenic purpura (TTP) in a subject comprising measuring a level of von Willebrand Factor (VWF) cleavage fragments in a blood sample from the subject before and after the treatment, wherein a decrease in amount of ultra-large multimer forms of VWF with a reduced triplet structure after the treatment indicates that the treatment is effective in increasing a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) activity or concentration in the subject.

### Claim 8 (depends on 7)

The method of claim 7, wherein measuring the level of VWF cleavage fragments comprises performing Western blot analysis with a VWF antibody to visualize VWF cleavage fragments.

### Claim 9 (depends on 8)

The method of claim 8, wherein the Western blot analysis is carried out under non-reducing conditions.

### Claim 10 (depends on 8)

The method of claim 8, wherein VWF fragments are visualized through use of a VWF antibody conjugated to a marker.

### Claim 11 (depends on 10)

The method of claim 10, wherein the marker is alkaline phosphatase (ALP) or horseradish peroxidase (HRP).

### Claim 12 (depends on 10)

The method of claim 10, wherein the marker is detected with enhanced chemiluminescence (ECL).

### Claim 13 (depends on 8)

The method of claim 8, wherein the VWF antibody is monoclonal.

### Claim 14 (depends on 7)

The method of claim 7, wherein VWF multimers are visualized by using high resolution horizontal SDS-agarose gel electrophoresis followed by immunostaining with a polyclonal rabbit anti-human VWF antibody.

### Claim 15 (depends on 7)

The method of claim 7, wherein the level of VWF cleavage fragments is detected at a sensitivity level of about 0.025 to about 0.05 Ag U/mL VWF.

### Claim 16

(canceled)

### Claim 17 (depends on 7)

The method of claim 7, wherein the blood sample is plasma or serum.

### Claim 18 (depends on 17)

The method of claim 17, wherein the blood sample is plasma.

### Claim 19 (depends on 7)

The method of claim 7, wherein the subject is a mammal.

### Claim at position 15

20-21. (canceled)

### Claim 22 (depends on 21)

The method of claim 21, wherein the subject is human.

### Claim at position 17

23-24. (canceled)

### Claim 25 (depends on 7)

The method of claim 7, wherein a change in VWF cleavage fragment level is detected by measuring the level of one or more of a 140 kDa VWF fragment or a 176 kDa VWF fragment.

### Claim 26 (depends on 25)

The method of claim 25, wherein the VWF fragment is a 176 kDa VWF fragment.

### Claim at position 20

27-42. (canceled)