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-9541549-B2
Application number	US-201414324620-A
Country	US
Kind code	B2
Filing date	Jul 7, 2014
Priority date	Apr 10, 2003
Publication date	Jan 10, 2017
Grant date	Jan 10, 2017

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 the determination of the mid-regional partial peptide of proadrenomedullin (mid-proAM) in a patient employing at least two antibodies capable of binding within the region of amino acids 60-92 of preproadrenomedullin (SEQ ID NO:1),
wherein the method comprises conducting a two-sided immunoassay by contacting plasma of the patient with two antibodies that specifically bind to different partial sequences of mid-proAM (SEQ ID NO: 3), so as to form sandwich complexes comprising the two antibodies and mid-proAM; and
detecting the sandwich complexes, thereby determining mid-proAM;
wherein one of the antibodies is obtained by immunization of an animal with an antigen which contains a synthetic peptide sequence which consists of 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 SEQ ID NO: 5.

2. A method according to claim 1 , wherein both antibodies are affinity-purified polyclonal antibodies.

3. A method according to claim 1 , wherein both antibodies are monoclonal antibodies.

4. A method according to claim 1 , wherein one of the antibodies is labeled and the other antibody is bound to a solid phase.

5. A method according to claim 1 , wherein both antibodies are present for the immunoassay dispersed in a liquid reaction mixture and that a first labeling component, which is part of a labeling system based on fluorescence or chemiluminescence extinction or amplification , is bound to a first antibody, and that a second labeling component of this labeling system is bound to a second antibody so that, after binding of both antibodies to the sequences, a measurable signal which permits detection of the resulting sandwich complexes in the immunoassay is generated.

Assignees

• Brahms Gmbh
• Brahms Gmbh

Inventors

• Bergmann Andreas
• Struck Joachim

Classifications

• G01N33/575 Primary

  for cancer · CPC title

• G01N33/574

  Physics · mapped topic

• G01N33/6893

  related to diseases not provided for elsewhere · CPC title

• G01N2800/26

  Infectious diseases, e.g. generalised sepsis · 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