Immunoassay using at least two pegylated analyte-specific binding agents

Patent metadata

Field	Value
Publication number	US-11099180-B2
Application number	US-201916528710-A
Country	US
Kind code	B2
Filing date	Aug 1, 2019
Priority date	Feb 2, 2017
Publication date	Aug 24, 2021
Grant date	Aug 24, 2021

Abstract

The disclosure concerns a method and kits for measurement of an analyte in a microparticle-based analyte-specific binding assay. In the assay, the microparticles are coated with the first partner of a binding pair, mixing the coated microparticles and at least two analyte-specific binding agents, each conjugated to the second partner of the binding pair, and a sample suspected of containing the analyte. The second partner of the binding pair is bound to each of the analyte-specific binding agents via a linker comprising from 12 to 30 ethylene glycol units (PEG 12 to 30), thereby binding the analyte via the conjugated analyte-specific binding agents to the coated microparticles. The method also entails separating the microparticles having the analyte bound via the binding pair and the analyte-specific binding agent from the mixture and measuring the analyte bound to the microparticles.

First claim

The invention claimed is:

1. A method for measurement of an analyte in a microparticle-based analyte-specific binding assay, wherein said microparticles are coated with the first partner of a binding pair, said method comprising
a) mixing the coated microparticles, at least two analyte-specific binding agents, wherein each of said analyte-specific binding agents is bound to a second partner of the binding pair, and a sample suspected of comprising or comprising the analyte, wherein said second partner of the binding pair is bound to each of said analyte-specific binding agents via a linker comprising from 12 to 30 ethylene glycol units (PEG 12 to 30), thereby binding the analyte via the said analyte-specific binding agents to the coated microparticles,
b) separating the microparticles comprising the analyte bound via the binding pair and the analyte-specific binding agent from the mixture and
c) measuring the analyte bound to the microparticles.

2. The method of claim 1 , wherein said measuring of the analyte bound to the microparticles is based on use of an electrochemiluminescent label.

3. The method of claim 1 , wherein said analyte comprises several variants.

4. The method of claim 1 , wherein said several variants are different genotypes, isoenzymes, isoforms, serotypes or mutants of said analyte.

5. The method of claim 1 , wherein said analyte is an antigen of an infectious agent.

6. The method of claim 1 , wherein said analyte is a viral antigen.

7. The method of claim 1 wherein said analyte is a hepatitis virus antigen or a human retroviral antigen.

8. The method of claim 1 , wherein said analyte is a hepatitis C virus or a hepatitis B virus or an HIV antigen.

9. The method of claim 1 , wherein said analyte is hepatitis C virus core antigen.

10. The method of claim 1 , wherein one of said at least two analyte-specific binding agents is an antibody binding to an epitope within the amino acid positions 140 to 172 of SEQ ID NO:1 and one of said at least two analyte-specific binding agents is an antibody binding to an epitope within the amino acid positions 20 to 80 of SEQ ID NO: 1.

11. A kit comprising in separate containers or in separated compartments of a single container unit at least microparticles coated with a first partner of a binding pair and at least two analyte-specific binding agents, each bound to the second partner of said binding pair, wherein said second partner of the binding pair is bound to each of said analyte-specific binding agents via a linker comprising from 12 to 30 ethylene glycol units (PEG 12 to 30).

12. The kit of claim 11 , wherein said first partner of a binding pair is avidin or streptavidin, and wherein said second partner of said binding pair is selected from biotin or biotin analogues.

13. The kit of claim 11 , wherein said at least two analyte-specific binding agents are viral antigen-specific binding agents.

14. The kit of claim 13 wherein one of said at least two viral antigen-specific binding agents is an antibody binding to an epitope within the amino acid positions 140 to 172 of SEQ ID NO:1.

15. The kit of claim 11 , further comprising in a separate container or in a separated compartment of a single container unit a further analyte-specific binding agent which is detectably labeled wherein said further analyte-specific binding agent which is detectably labeled is an antibody binding to an epitope within amino acid positions 100-120 of SEQ ID NO:1.

Assignees

• Roche Diagnostics Operations Inc

Inventors

• Bollhagen Ralf
• Upmeier Barbara
• Naser Werner
• Zarnt Toralf

Classifications

• G01N33/5767

  non-A, non-B hepatitis · CPC title

• G01N33/56983

  Viruses · CPC title

• G01N33/54313 Primary

  the carrier being characterised by its particulate form · CPC title

• G01N2333/186

  Hepatitis C; Hepatitis NANB · CPC title

• G01N33/56988 Primary

  HIV or HTLV · CPC title