Determination of a midregional proadrenomedullin partial peptide in biological fluids for diagnostic purposes, and immunoassays for carrying out such a determination

Patent metadata

Field	Value
Publication number	US-9885709-B2
Application number	US-55129804-A
Country	US
Kind code	B2
Filing date	Jan 29, 2004
Priority date	Apr 10, 2003
Publication date	Feb 6, 2018
Grant date	Feb 6, 2018

Abstract

Method for the determination of adrenomedullin immunoreactivity in biological fluids for diagnostic purposes, in particular in sepsis, cardiac and cancer diagnosis, in which the midregional partial peptide (mid-proAM; SEQ ID NO:3) of proadrenomedullin, which comprises the amino acids (45-92) of the complete preproadrenomedullin (pre-proAM; SEQ ID NO:1), is measured in particular by means of an immunoassay which operates with at least one labelled antibody which specifically recognizes a sequence of mid-proAM.

First claim

The invention claimed is:

1. A method for detecting and quantitating in a biological fluid sample from a human the mid-regional partial peptide of proadrenomedullin (mid-proAM) which consists of the sequence of SEQ ID NO: 3, comprising
(a) contacting the sample with a labeled monoclonal or polyclonal antibody which specifically binds to said mid-proAM partial peptide, and
(b) detecting and quantitating the resulting peptide:antibody complex using an immunoassay, wherein said immunoassay
(i) is not a radioimmunoassay, and
(ii) has a limit of detection of about 50 pmol/l; and

wherein said immunoassay using said at least one labeled antibody is a sandwich assay further employing at least one additional antibody which specifically binds to a different partial sequence of mid-proAM (SEQ ID NO: 3) from that bound by said at least one labeled antibody; and wherein for said sandwich assay, one of the antibodies is obtained by immunization of an animal with an antigen which contains a synthetic peptide sequence which consists of amino acids 69-86 of pre-proAM (SEQ ID NO: 4), and the other of the antibodies is obtained by immunization with an antigen which contains a synthetic peptide sequence which consists of amino acids 83-94 of pre-proAM (SEQ ID NO: 5).

2. An immunoassay method for detecting and quantitating in a biological fluid sample the mid-regional partial peptide of proadrenomedullin (mid-proAM) which consists of the sequence of SEQ ID NO: 3, comprising contacting the sample with at least one labeled monoclonal or polyclonal antibody which specifically binds to said mid-proAM partial peptide, and detecting the thus-formed antibody:mid-proAM complex,
wherein said immunoassay
(a) is not a radioimmunoassay, and
(b) has a limit of detection of about 50 pmol/l;

said immunoassay being a sandwich assay further employing at least one additional antibody which specifically binds to a different partial sequence of mid-proAM (SEQ ID NO: 3) from that bound by said at least one labeled antibody; and wherein for said sandwich assay, one of the antibodies is obtained by immunization of an animal with an antigen which contains a synthetic peptide sequence which consists of amino acids 69-86 of pre-proAM (SEQ ID NO: 4), and the other of the antibodies is obtained by immunization with an antigen which contains a synthetic peptide sequence which consists of amino acids 83-94 of pre-proAM (SEQ ID NO: 5).

Assignees

• Bergmann Andreas
• Struck Joachim
• Brahms Gmbh

Inventors

• Bergmann Andreas
• Struck Joachim

Classifications

• G01N33/575 Primary

  for cancer · CPC title

• G01N33/574

  Physics · mapped topic

• G01N33/54306 Primary

  Solid-phase reaction mechanisms · CPC title

• G01N33/6893

  related to diseases not provided for elsewhere · CPC title

• G01N33/74

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