ASSAYS FOR DETECTING THE PRESENCE OR AMOUNT OF AN ANTI-DRUG ANTIBODY

• US 20150226758A1
• Filed: 02/11/2015
• Published: 08/13/2015
• Est. Priority Date: 02/11/2014
• Status: Active Grant

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Grabert, Ryan, Richards, Susan, Theobald, Valerie, Xu, Yuanxin, Zoghbi, Jad
• Granted Patent: US 9,759,732 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: G01N 2430/00 Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/539 involving precipitating reagent , e.g. ammonium sulfate
• CPC Class Codes: G01N 33/6854 Immunoglobulins
• CPC Class Codes: G01N 33/94 involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors

Abstract

Methods and kits for detecting antibodies (e.g., anti-drug antibodies). Such methods and kits permit the detection of, for example, anti-drug antibodies in human body fluids, such as blood, plasma and serum.

Claims

1. A method for determining the presence or absence of an anti-drug antibody (ADA) in a sample, the method comprising:

contacting the sample with an excess amount of drug to which the ADA binds to form drug/ADA complexes;

contacting the drug/ADA complexes with polyethylene glycol (PEG), to form a precipitate comprising drug/ADA complexes;

contacting the precipitate with a solution, dissociate the drug/ADA complexes;

immobilizing the dissociated ADAs on a substrate; and

determining if the ADA is present or absent in the sample.

2. The method of claim 1, wherein the determining step comprises

contacting the immobilized ADA with drug labeled with a detectable label;

and

determining the presence or absence of said detectable label, to thereby determine the presence or absence of ADA in the sample.

3. The method of claim 1, wherein the substrate is a high bind carbon plate.

4. The method of claim 1, wherein the solution to dissociate the drug/ADA complexes comprises an acid.

5. The method of claim 1, wherein the solution to dissociate the drug/ADA complexes comprises a base.

6. The method of claim 1, wherein the substrate comprises a carbon surface, glass surface, silica surface, metal surface, a polymeric material, a surface containing a metallic or chemical coating, a membrane, a bead, a porous polymer matrix, or substrates comprising cellulosic fibers, or any combination thereof.

7. The method of claim 2, wherein the detectable label comprises a label selected from the group consisting of a radioactive isotope, an enzyme, a fluorescent label, a chemiluminescent label, and electrochemiluminescent label, and a substrate for an enzymatic detection reaction.

8. The method of claim 1, wherein the drug comprises an antibody or fragment thereof, nucleic acid, peptide, polypeptide, peptidomimetic, carbohydrate, lipid, or an organic or inorganic small molecule compound, or any combination thereof.

9. The method of claim 1, wherein the drug is a drug modified to exhibit less immunogenicity as compared to the same drug in unmodified form.

10. The method of claim 1, wherein the PEG comprises at least one PEG selected from the group consisting of PEG1000, PEG1450, PEG3000, PEG6000, PEG8000, PEG10000, PEG14000, PEG15000, PEG20000, PEG250000, PEG30000, PEG35000, and PEG40000.

11. The method of claim 1, wherein the sample is contacted with PEG at a concentration of between about 0.1% and about 10.0%,

12. The method of claim 4, wherein the acid is at a concentration of between about 0.1M to about 5M.

13. The method claim 12, wherein the acid is selected from the group consisting of citric acid, isocitric acid, glutamic acid, acetic acid, lactic acid, formic acid, oxalic acid, uric acid, trifluoroacetic acid, benzene sulfonic acid, aminomethanesulfonic acid, camphor-10-sulfonic acid, chloroacetic acid, bromoacetic acid, iodoacetic acid, propanoic acid, butanoic acid, glyceric acid, succinic acid, malic acid, aspartic acid, hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, boric acid, hydrofluoric acid, hydrobromic acid, and any combination thereof.

14. The method of claim 1, wherein the sample comprises the drug.

15. The method of claim 1, wherein the method further comprises immobilizing the drug on the substrate before or after the step of immobilizing the ADA on the substrate.

16. The method of claim 1, wherein the sample comprises a biological sample, wherein the biological sample comprises material selected from the group consisting of body fluids, mucus secretions, saliva, blood, whole blood, plasma, and serum.

17. A method for reducing interference in a drug assay due to the presence of an anti-drug antibody (ADA) in a sample, the method comprising:

contacting the sample with an excess amount of an ADA to saturate free drug and form drug/ADA complexes;

contacting the drug/ADA complexes with polyethylene glycol (PEG), to thereby form a precipitate comprising drug/ADA complexes;

contacting the precipitate with a solution, to dissociate the drug/ADA complexes;

immobilizing the dissociated drug on a substrate; and

performing the drug assay, to thereby reduce interference from the ADA.

18. The method of claim 17, wherein the drug assay comprises a drug quantitation assay, a drug PK assay or a drug potency assay, or combination thereof.

19. The method of claim 18, wherein the drug comprises an antibody or fragment thereof, nucleic acid, peptide, polypeptide, peptidomimetic, carbohydrate, lipid, or an organic or inorganic small molecule compound.

20. The method of claim 19, further comprising determining the presence or absence of the drug in the sample using an anti-idiotype antibody labeled with a detectable label.