Method for the detection of a binding partner of a multispecific binder

Patent metadata

Field	Value
Publication number	US-9945869-B2
Application number	US-201414449437-A
Country	US
Kind code	B2
Filing date	Aug 1, 2014
Priority date	Feb 1, 2012
Publication date	Apr 17, 2018
Grant date	Apr 17, 2018

Abstract

Herein is reported a method for the detection of free antigen of a multispecific antibody in a sample, whereby the antigen to be detected can be specifically bound by a first binding site of the multispecific antibody, comprising the step of incubating a sample comprising free antigen and multispecific antibody with an anti-idiotypic antibody, which specifically binds to a second binding specificity of the bispecific antibody, which is different from the first binding specificity, whereby the anti-idiotypic antibody is bound to a solid phase.

First claim

The invention claimed is:

1. A method for determining the presence and/or the amount of an antigen of a bispecific antibody in a sample, wherein the antigen to be detected can be specifically bound by a first binding specificity of the bispecific antibody, and wherein the antigen is complexed to the bispecific antibody (antigen-bispecific antibody-complex), comprising
incubating the sample comprising the antigen and the bispecific antibody with an anti-idiotypic antibody, which specifically binds to a second binding specificity of the bispecific antibody, which is different from the first binding specificity, wherein the anti-idiotypic antibody is bound to a solid phase, under conditions where an antigen-bispecific antibody-anti-idiotypic antibody complex can be formed; and
incubating the antigen-bispecific antibody-anti-idiotypic antibody complex with an antibody that specifically binds to the antigen at an epitope different from the epitope bound by the bispecific antibody;
and determining the presence and/or the amount of the antigen of the bispecific antibody which is complexed to the bispecific antibody in a sample.

2. The method according to claim 1 , further comprising the step of:
detecting the antigen-bispecific antibody-anti-idiotypic antibody complex and thereby determining the presence and/or the amount of the antigen of the bispecific antibody.

3. The method according to claim 1 comprising the following steps:
providing the sample comprising the antigen and the bispecific antibody, wherein at least 90% of the antigen are complexed by the bispecific antibody in the antigen-bispecific antibody complex, wherein the antigen to be detected can be specifically bound by the first binding specificity of the bispecific antibody;
incubating the sample comprising the antigen and the bispecific antibody with the anti-diotypic antibody, which specifically binds to the second binding specificity of the bispecific antibody, which is different from the first binding specificity, wherein the anti-idiotypic antibody is bound to the solid phase; and
incubating the antigen-bispecific antibody complex with the antibody that specifically binds to the antigen at an epitope different from the epitope bound by the bispecific antibody,
and thereby determining the presence and/or the amount of the antigen of the bispecific antibody in the sample.

4. The method according to claim 1 , comprising the following steps:
incubating the sample comprising the antigen and the bispecific antibody with an amount of the bispecific antibody to provide a sample wherein at least 90% of the antigen is complexed by the bispecific antibody in the antigen-bispecific antibody complex, wherein the antigen to be detected can be specifically bound by the first binding specificity of the bispecific antibody;
incubating the sample comprising the antigen-bispecific antibody complex with the anti-idiotypic antibody, which specifically binds to the second binding specificity of the bispecific antibody, which is different from the first binding specificity, wherein the anti-idiotypic antibody is bound to the solid phase, and wherein the antigen-bispecific antibody-anti-idiotypic antibody complex can be formed; and
incubating the antigen-bispecific antibody-anti-idiotypic antibody complex with the antibody that specifically binds to the antigen at an epitope different from the epitope bound by the bispecific antibody,
and thereby determining the presence and/or the amount of the antigen of the bispecific antibody in the sample.

5. The method according to claim 4 , wherein the amount of the bispecific antibody is between 1 μg/ml and 10 μg/ml sample.

6. The method according to claim 4 , wherein at least 95% of the antigen is complexed by the bispecific antibody.

7. The method according to claim 6 , wherein at least 98% of the antigen is complexed by the bispecific antibody.

8. A method for determining the amount of an antigen of a bispecific antibody that is complexed to the bispecific antibody in a sample comprising:
incubating a sample comprising the antigen and the bispecific antibody with an anti-idiotypic antibody, which specifically binds to a binding specificity of the bispecific antibody, which is different from the binding specificity by which the antigen is bound, wherein an antigen-bispecific antibody-anti-idiotypic antibody complex can be formed, wherein the anti-idiotypic antibody is bound to a solid phase; and
incubating the antigen-bispecific antibody-anti-idiotypic antibody complex with an antibody that specifically binds to the antigen at an epitope that is different from the epitope bound by the bispecific antibody,
and thereby determining the amount of an antigen of a bispecific antibody complexed to the bispecific antibody in a sample.

Assignees

• Hoffmann La Roche

Inventors

• Stubenrauch Kay-Gunnar
• Wessels Uwe

Classifications

• G01N2333/00

  Assays involving biological materials from specific organisms or of a specific nature · CPC title

• G01N33/54306 Primary

  Solid-phase reaction mechanisms · CPC title

• G01N33/543 Primary

  with an insoluble carrier for immobilising immunochemicals · CPC title

• G01N33/6854 Primary

  Immunoglobulins · CPC title