BACKGROUND: Adrenomedullin (ADM) is a circulating peptide known to regulate vasodilation and vascular integrity. Increased plasma ADM concentrations have been described for several life-threatening conditions, including cardiovascular diseases and septic shock. Reliable methods for the simple quantification of bioactive ADM (bio-ADM) are lacking. METHODS: Monoclonal antibodies against the amidated C-terminus and middle portion of bio-ADM were generated and used for the development of a 1-step immunometric assay for the specific quantification of bio-ADM in plasma. The assay was developed in a microtiter plate/chemiluminescence label format with a significantly reduced incubation time. Precision, linearity, specimen stability, and distribution of results in healthy subjects were evaluated. RESULTS: The use of monoclonal antibodies against predetermined epitopes of bio-ADM enabled the development of an assay for the determination of bio-ADM directly in EDTA plasma. Plasma samples were stable for up to 24 h at ambient temperature and over multiple freeze-thaw cycles without loss of immunoreactivity. The assay had a limit of detection of 3 pg/mL and a limit of quantification of 11 pg/mL. The assay exhibited acceptable linearity characteristics and was not influenced by complement factor H, a putative ADM-binding protein. In healthy subjects, bio-ADM concentrations were all above the limit of detection, and approximately half of them were above the limit of quantification. CONCLUSIONS: By using monoclonal antibodies with defined epitope specificities, we have developed a simple, rapid, accurate, and sensitive sandwich immunoassay for bio-ADM. The assay is a potentially novel tool to support patient management, particularly in acute care in the field of sepsis and other indications, which are currently being investigated, such as acute heart failure.
BACKGROUND:Adrenomedullin (ADM) is a circulating peptide known to regulate vasodilation and vascular integrity. Increased plasma ADM concentrations have been described for several life-threatening conditions, including cardiovascular diseases and septic shock. Reliable methods for the simple quantification of bioactive ADM (bio-ADM) are lacking. METHODS: Monoclonal antibodies against the amidated C-terminus and middle portion of bio-ADM were generated and used for the development of a 1-step immunometric assay for the specific quantification of bio-ADM in plasma. The assay was developed in a microtiter plate/chemiluminescence label format with a significantly reduced incubation time. Precision, linearity, specimen stability, and distribution of results in healthy subjects were evaluated. RESULTS: The use of monoclonal antibodies against predetermined epitopes of bio-ADM enabled the development of an assay for the determination of bio-ADM directly in EDTA plasma. Plasma samples were stable for up to 24 h at ambient temperature and over multiple freeze-thaw cycles without loss of immunoreactivity. The assay had a limit of detection of 3 pg/mL and a limit of quantification of 11 pg/mL. The assay exhibited acceptable linearity characteristics and was not influenced by complement factor H, a putative ADM-binding protein. In healthy subjects, bio-ADM concentrations were all above the limit of detection, and approximately half of them were above the limit of quantification. CONCLUSIONS: By using monoclonal antibodies with defined epitope specificities, we have developed a simple, rapid, accurate, and sensitive sandwich immunoassay for bio-ADM. The assay is a potentially novel tool to support patient management, particularly in acute care in the field of sepsis and other indications, which are currently being investigated, such as acute heart failure.
Authors: Wouter C Meijers; Antoni Bayes-Genis; Alexandre Mebazaa; Johann Bauersachs; John G F Cleland; Andrew J S Coats; James L Januzzi; Alan S Maisel; Kenneth McDonald; Thomas Mueller; A Mark Richards; Petar Seferovic; Christian Mueller; Rudolf A de Boer Journal: Eur J Heart Fail Date: 2021-10-10 Impact factor: 17.349
Authors: Oscar H M Lundberg; Mari Rosenqvist; Kevin Bronton; Janin Schulte; Hans Friberg; Olle Melander Journal: PLoS One Date: 2022-04-28 Impact factor: 3.752
Authors: Jacqueline Lammert; Maryam Basrai; Joachim Struck; Oliver Hartmann; Christoph Engel; Stephan C Bischoff; Anika Berling-Ernst; Martin Halle; Marion Kiechle; Sabine Grill Journal: Geburtshilfe Frauenheilkd Date: 2022-06-03 Impact factor: 2.754
Authors: Oscar H M Lundberg; Maria Lengquist; Martin Spångfors; Martin Annborn; Deborah Bergmann; Janin Schulte; Helena Levin; Olle Melander; Attila Frigyesi; Hans Friberg Journal: Crit Care Date: 2020-11-04 Impact factor: 9.097