Specific Detection of Rat Antibodies in Mouse Serum

Patent metadata

Field	Value
Publication number	US-2016266135-A2
Application number	US-201514858449-A
Country	US
Kind code	A2
Filing date	Sep 18, 2015
Priority date	Mar 20, 2013
Publication date	Sep 15, 2016
Grant date	—

Abstract

One aspect as reported herein is a method for detecting a rat antibody in a serum or plasma sample (obtained) from a mouse comprising the steps of a) providing the sample to be analyzed, b) incubating said serum or plasma sample with an antibody that specifically binds to rat IgG and that does not specifically bind to mouse IgG, wherein the antibody is i) a mixture of an antibody binding to rat kappa light chain and an antibody binding to rat lambda light chain, or ii) a mixture of an antibody binding to rat IgG1 with an avidity of 4.1×1010 M−1 or more, an antibody binding to rat IgG2a with an avidity of 8.6×109 M−1 or more, an antibody binding to rat IgG2b with an avidity of 6.4×1010 M−1 or more and an antibody binding to rat IgG2c with an avidity of 9.5×1010 M−1 or more, c) optionally incubating said sample with a reagent appropriate for the selective detection of total, active or antigen-bound rat antibody, and d) correlating the complex formed in (b) or (c) to the concentration of the rat antibody in the sample.

First claim

What is claimed is:

1 . Method for detecting a rat antibody in a serum or plasma sample obtained from a mouse comprising the steps of
a) providing the sample to be analyzed, b) incubating said serum or plasma sample with an antibody that specifically binds to rat IgG and that does not specifically bind to mouse IgG, wherein the antibody is
i) a mixture of a monoclonal antibody binding to rat kappa light chain and a monoclonal antibody binding to rat lambda light chain, or
ii) a mixture of a monoclonal antibody binding to rat IgG1 with an avidity of 4.1×10 10 M −1 or more, a monoclonal antibody binding to rat IgG2a with an avidity of 8.6×10 9 M −1 or more, a monoclonal antibody binding to rat IgG2b with an avidity of 6.4×10 10 M −1 or more and a monoclonal antibody binding to rat IgG2c with an avidity of 9.5×10 10 M −1 or more, and
c) correlating the complex formed in (b) to the concentration of the rat antibody in the sample.

2 . Method for detecting a rat antibody in a serum or plasma sample obtained from a mouse comprising the steps of
a) incubating said serum or plasma sample with a capture antibody conjugated to a solid phase and thereby forming a capture antibody-rat antibody complex, b) incubating the complex formed in (a) with a tracer antibody and thereby correlating the complex formed in (a) to the presence or concentration of said rat antibody,
wherein the capture antibody specifically binds to rat IgG and that does not specifically bind to mouse IgG and is
i) a mixture of a monoclonal antibody binding to rat kappa light chain and a monoclonal antibody binding to rat lambda light chain, or
ii) a mixture of a monoclonal antibody binding to rat IgG1 with an avidity of 4.1×10 10 M −1 or more, a monoclonal antibody binding to rat IgG2a with an avidity of 8.6×10 9 M −1 or more, a monoclonal antibody binding to rat IgG2b with an avidity of 6.4×10 10 M −t or more and a monoclonal antibody binding to rat IgG2c with an avidity of 9.5×10 10 M −1 or more.

3 . Method for detecting a rat antibody in a serum or plasma sample obtained from a mouse comprising the steps of
a) incubating said serum or plasma sample with a capture antibody conjugated to a solid phase and thereby forming a capture antibody-rat antibody complex, b) incubating the complex formed in (a) with a tracer antibody and thereby correlating the complex formed in (a) to the presence or concentration of said rat antibody,
wherein the tracer antibody specifically binds to rat IgG and that does not specifically bind to mouse IgG and is
i) a mixture of a monoclonal antibody binding to rat kappa light chain and an antibody binding to rat lambda light chain, or
ii) a mixture of a monoclonal antibody binding to rat IgG1 with an avidity of 4.1×10 10 M −1 or more, a monoclonal antibody binding to rat IgG2a with an avidity of 8.6×10 9 M −1 or more, a monoclonal antibody binding to rat IgG2b with an avidity of 6.4×10 10 M −1 or more and a monoclonal antibody binding to rat IgG2c with an avidity of 9.5×10 10 M −1 or more.

4 - 5 . (canceled)

6 . Method according to claim 2 , characterized in that the tracer antibody and the capture antibody are independently of each other
i) a mixture of a monoclonal antibody binding to rat kappa light chain and a monoclonal antibody binding to rat lambda light chain, or ii) a mixture of a monoclonal antibody binding to rat IgG1 with an avidity of 4.1×10 10 M −1 or more, a monoclonal antibody binding to rat IgG2a with an avidity of 8.6×10 9 M −1 or more, a monoclonal antibody binding to rat IgG2b with an avidity of 6.4×10 1 ° M −1 or more and a monoclonal antibody binding to rat IgG2c with an avidity of 9.5×10 10 M −1 or more.

7 . Method according to any one of claims 1 to 3 , characterized in that the method is an antigen bridging immunoassay.

8 . Method according to any one of claims 1 to 3 , characterized in that the method is a sandwich immunoassay.

9 . Method according to claim 2 or 3 , characterized in that the capture antibody is conjugated to biotin and immobilization is performed via immobilized avidin or streptavidin.

10 . Method according to claim 2 or 3 , characterized in that the tracer antibody is conjugated to digoxygenin and linking to the detectable label is performed via an antibody against digoxygenin.

11 . The method according to any one of claims 1 to 3 , characterized in that said rat antibody is a Fab.

12 - 14 . (canceled)

Assignees

• Hoffmann La Roche

Inventors

• Stubenrauch Kay-Gunnar
• Wessels Uwe

Classifications

• G01N33/94

  involving narcotics {or drugs or pharmaceuticals, neurotransmitters or associated receptors} · CPC title

• G01N33/686 Primary

  Anti-idiotype · CPC title

• G01N33/6854 Primary

  Immunoglobulins · CPC title

• G01N33/577

  involving monoclonal antibodies {binding reaction mechanisms characterised by the use of monoclonal antibodies (G01N33/5302 - G01N33/576 take precedence)} · CPC title

• G01N33/6803

  General methods of protein analysis not limited to specific proteins or families of proteins · CPC title