Competitive ligand binding assay for detecting neutralizing antibodies

Patent metadata

Field	Value
Publication number	US-2016252520-A1
Application number	US-201415031126-A
Country	US
Kind code	A1
Filing date	Oct 30, 2014
Priority date	Oct 31, 2013
Publication date	Sep 1, 2016
Grant date	—

Abstract

A immunogenicity assay for detecting the presence of neutralizing antibodies to a biotherapeutic protein wherein the biotherapeutic protein has been administered to a patient in need, comprising the steps of (a) obtaining a sample from the patient; (b) incubating the sample in the presence of a capture reagent; and (c) adding a detecting reagent, wherein a decreased signal relative to a control sample indicates the presence of a neutralizing antibody to the biotherapeutic agent.

First claim

What is claimed is:

1 . A method for detecting the presence of neutralizing antibodies to a biotherapeutic protein wherein the biotherapeutic protein has been administered to a patient in need, comprising the steps of (a) combining a patient sample with a capture reagent, and (b) adding a detecting reagent, wherein a decreased signal relative to a control sample indicates the presence of a neutralizing antibody to the biotherapeutic agent.

2 . The method of claim 1 , wherein the protein biotherapeutic is a monoclonal antibody

3 . The method of claim 1 or 2 , wherein the protein biotherapeutic is an anti-interleukin-6 receptor α (IL-6Rα) monoclonal antibody.

4 . The method of any one of claim 1 - 3 , wherein the protein biotherapeutic is sarilumab or tocilizumab.

5 . The method of claim 4 , wherein the protein biotherapeutic is sarilumab.

6 . The method of claim 5 , wherein the patient is treated for an IL-6-dependent disease.

7 . The method of claim 6 , wherein the IL-6-dependent disease is one of rheumatoid arthritis, diabetes, atherosclerosis, Alheimer's disease, systemic lupus erythematosus, multiple myeloma, connective tissue disorders, Castleman's disease and prostate cancer.

8 . The method of claim 7 , wherein the IL-6-dependent disease is rheumatoid arthritis.

9 . The method of any one of the above claims, wherein the capture reagent comprises the labeled biotherapeutic protein.

10 . The method of any one of the above claims, wherein the capture reagent is labeled sarilumab.

11 . The method of claim 10 , wherein the labeled sarilumab is biotinylated sarilumab.

12 . The method of claim 11 , wherein the biotinylated sarilumab is bound to an avidin-coated plate.

13 . The method of any of the above claims, wherein the detection reagent is labeled soluble IL-6 receptor α.

14 . The method of claim 13 , wherein the detection reagent is ruthenium labeled soluble IL-6 receptor α.

15 . The method of any of the above claims wherein the patient sample is subjected to treatment with low pH prior to step (b).

16 . The method of claim 15 , wherein the patient sample is subjected to treatment with acetic acid prior to step (b).

17 . The method of claim 15 or 16 , wherein the treatment with low pH is followed by neutralization.

18 . The method of any of the above claims, wherein the assay exhibits a sensitivity tolerance of about 150 ng/mL, a drug tolerance of about 500 ng/mL, and a target interference tolerance of about 1 μg/mL.

19 . A method for detecting the presence of neutralizing antibodies to sarilumab, wherein sarilumab has been administered to a patient in need, comprising the steps of (a) combining a patient serum sample with a capture reagent comprising biotinylated sarilumab, (b) subjecting the combined serum sample and capture reagent to treatment with low pH, followed by a neutralizing step; and (d) adding a detecting reagent, wherein the detecting reagent comprises labeled soluble IL-6 receptor α, wherein a decreased signal relative to a control sample indicates the presence of a neutralizing antibody to the biotherapeutic agent.

20 . The method of claim 19 , wherein the detection reagent is ruthenium labeled soluble IL-6 receptor α.

21 . The method of claims 19 - 20 , wherein the patient is suffering from rheumatoid arthritis.

22 . The method of any of claims 19 - 21 , wherein the assay exhibits a sensitivity tolerance of about 150 ng/mL, a drug tolerance of about 500 ng/mL, and a target interference tolerance of about 1 μg/mL.

23 . A kit, comprising the capture and detection reagents of any of the above claims and instructions for use.

Assignees

• Regeneron Pharma

Inventors

• Rajadhyaksha Manoj
• Shyu Gary
• Georgaros Camille
• Partridge Michael
• Torri Albert

Classifications

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P37/02

  Immunomodulators · CPC title