One-Step Direct Assay for Mature-type Adrenomedullin with Monoclonal Antibodies | Zendy

Hideki Ohta | Zendy; Tetsuo Tsuji | Zendy; Shigeru Asai | Zendy; Kazuyuki Sasakura | Zendy; Hiroshi Teraoka | Zendy; Kazuo Kitamura | Zendy; Kenji Kangawa | Zendy

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One-Step Direct Assay for Mature-type Adrenomedullin with Monoclonal Antibodies

Author(s) -

Hideki Ohta,

Tetsuo Tsuji,

Shigeru Asai,

Kazuyuki Sasakura,

Hiroshi Teraoka,

Kazuo Kitamura,

Kenji Kangawa

Publication year - 1999

Publication title -

clinical chemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.705

H-Index - 218

eISSN - 1530-8561

pISSN - 0009-9147

DOI - 10.1093/clinchem/45.2.244

Subject(s) - adrenomedullin , monoclonal antibody , chromatography , chemistry , human plasma , monoclonal , detection limit , peptide , radioimmunoassay , antibody , extraction (chemistry) , dilution , medicine , biochemistry , immunology , physics , thermodynamics

Adrenomedullin (AM) is a potent hypotensive peptide. Plasma contains mature-type AM (m-AM), which is amidated at the carboxy terminus, and an intermediate, AM-Gly. We developed a one-step two-site IRMA specific for determining human m-AM with monoclonal antibodies. The detection limit was 0.5 pmol/L, and the working range (CV <15%) was 1-300 pmol/L. Dilution of plasma samples showed good linearity. The recovery of added AM was 91-118%. The intra- and interassay imprecision values (CVs) were 4.4-8.2% and 5.5-8.3%, respectively. The assay had no cross-reactivity with AM-Gly or other peptides similar to AM. The mean (+/- SD) plasma human m-AM concentration of 61 healthy subjects was 1.18 +/- 0.65 pmol/L. In conclusion, our IRMA makes it possible to specifically measure m-AM, using a small amount of plasma sample (0.2 mL) by a one-step overnight assay without prior extraction. Our simplified method would be suitable for clinical studies on AM, especially when large numbers of samples must be processed.

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