Augurin immunoassay

Patent metadata

Field	Value
Publication number	US-10024872-B2
Application number	US-201414903893-A
Country	US
Kind code	B2
Filing date	Jul 10, 2014
Priority date	Jul 12, 2013
Publication date	Jul 17, 2018
Grant date	Jul 17, 2018

Abstract

The present invention relates to an immunoassay method for the detection of augurin or a precursor or fragment thereof comprising contacting a sample suspected of comprising augurin or a precursor or fragment thereof with a first and second antibody specific for augurin or a precursor or fragment thereof, wherein said first and second antibodies or antigen-binding fragments or derivatives thereof are specific for epitopes comprised in the sequence spanning amino acids 71 to 107 of pre-augurin according to SEQ ID NO:1.

First claim

The invention claimed is:

1. An immunoassay method for the detection of augurin or a precursor or fragment thereof comprising
(a) contacting a sample suspected of comprising augurin or a precursor or fragment thereof with a first antibody or an antigen-binding fragment thereof specific for augurin or a precursor or fragment thereof and a second antibody or an antigen-binding fragment thereof specific for augurin or a precursor or fragment thereof under conditions allowing for the binding of the two antibodies or antigen-binding fragments thereof to augurin or a precursor or fragment thereof,
wherein said first and second antibodies or antigen-binding fragments thereof are specific for epitopes contained within the sequence spanning amino acids 71 to 107 of pre-augurin according to SEQ ID NO:1, wherein said first and second antibodies or antigen-binding fragments thereof are specific for different and non-overlapping epitopes,
wherein the first antibody or antigen-binding fragment is produced by a hybridoma cell line selected from the group consisting of cell line 482/H2 deposited as DSM ACC3208, cell line 482/H10 deposited as DSM ACC3209, cell line 482/H7 deposited as DSM ACC3210, and cell line 482/G9 deposited as DSM ACC3211, and
wherein the second antibody or antigen-binding fragment is produced by hybridoma cell line 439/F4 deposited as DSM ACC3206 or hybridoma cell line 439/H10 deposited as DSM ACC3207, and

(b) detecting the binding of the first and second antibodies or antigen-binding fragments thereof to augurin or a precursor or fragment thereof.

2. An immunoassay method for the detection of augurin or a precursor or fragment thereof comprising
(a) contacting a sample suspected of comprising augurin or a precursor or fragment thereof with a first antibody or an antigen-binding fragment thereof specific for augurin or a precursor or fragment thereof under conditions allowing for the formation of a complex between augurin or a precursor or fragment thereof and said first antibody or antigen-binding fragment thereof,
(b) contacting said sample with a second antibody or an antigen-binding fragment thereof specific for augurin or a precursor or fragment thereof under conditions allowing for the formation of a ternary complex between augurin or a precursor or fragment thereof and said first and second antibodies or antigen-binding fragments thereof,
wherein said first and second antibodies or antigen-binding fragments thereof are specific for epitopes contained within the sequence spanning amino acids 71 to 107 of pre-augurin according to SEQ ID NO:1,
wherein said first and second antibodies or antigen-binding fragments thereof are specific for different and non-overlapping epitopes,
wherein the first antibody or antigen-binding fragment is produced by a hybridoma cell line selected from the group consisting of cell line 482/H2 deposited as DSM ACC3208, cell line 482/H10 deposited as DSM ACC3209, cell line 482/H7 deposited as DSM ACC3210, and cell line 482/G9 deposited as DSM ACC3211, and
wherein the second antibody or antigen-binding fragment is produced by hybridoma cell line 439/F4 deposited as DSM ACC3206 or hybridoma cell line 439/H10 deposited as DSM ACC3207, and

(c) detecting said ternary complex.

3. The immunoassay method according to claim 1 , wherein said epitopes are not more than 6 amino acids apart.

4. The immunoassay method according to claim 3 , wherein said first and second antibodies or antigen-binding fragments thereof are specific for epitopes contained within the sequence spanning amino acids 71 to 88 according to SEQ ID NO:1 and wherein said epitopes are not more than 3 amino acids apart.

5. The immunoassay method according to claim 1 , wherein the first antibody or antigen-binding fragment thereof is specific for an epitope contained within the sequence spanning amino acids 71 to 83 of pre-augurin according to SEQ ID NO:1 and wherein the second antibody or antigen-binding fragment thereof is specific for an epitope contained within the sequence spanning amino acids 79 to 88 of pre-augurin according to SEQ ID NO:1.

6. The immunoassay method according to claim 1 , wherein the sample is a derived from a bodily fluid or tissue of a subject.

7. The immunoassay method according to claim 1 , wherein the immunoassay is a sandwich ELISA assay.

8. The immunoassay method according to claim 1 , wherein the first antibody or the antigen-binding fragment thereof is a solid phase antibody that is attached a solid phase and the second antibody or the antigen-binding fragment thereof is a tracer antibody labelled with a detection probe, or vice versa.

9. The immunoassay method according to claim 8 , wherein the solid phase is a bead, a surface of a well or a container, a chip, and/or a strip.

10. The immunoassay method according to claim 8 , wherein the detection probe is a dye, a radioisotope, and/or a reactive or catalytically active moiety.

Assignees

• Brahms Gmbh
• Brahms Gmbh

Inventors

• Struck Joachim
• Ziera Tim

Classifications

• G01N33/74 Primary

  involving hormones {or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors} · CPC title

• C12N5/12

  Fused cells, e.g. hybridomas · CPC title

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07K16/26

  against hormones {; against hormone releasing or inhibiting factors} · CPC title

• G01N2333/575

  Hormones · CPC title