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