Measurement of corticotropin in unextracted plasma: comparison of a time-resolved immunofluorometric assay and an immunoradiometric assay, with use of the same monoclonal antibodies.
We describe a time-resolved immunofluorometric assay (IFMA) for corticotropin in unextracted human plasma, based on the use of two monoclonal antibodies: europium-labeled antibody 1A12 and antibody 2A3 coated onto microtiter wells. We compared the results of this assay with those of an immunoradiometric assay (IRMA) performed with the same antibodies working ranges (CV less than 10%) were 25 to 1000 ng/L and 22 to 1000 ng/L for the IFMA and IRMA, respectively, and both assays had comparable detection limits (IFMA 4.0 +/- 1 ng/L, IRMA 3.5 +/- 0.8 ng/L). Results by both assays for 130 patients' samples containing corticotropin within the range 3-100 ng/L and greater than ng/L correlated well (r = 0.88 and 0.92, respectively), and samples with corticotropin in the range 80-624 ng/L gave results that paralleled those for the standard curve. Corticotropin concentrations in apparently healthy subjects were consistent with those reported previously. The IFMA is a simple, precise, and robust assay that can be completed within one day. Its nonisotopic label is stable for at least 50 weeks.
Dobson
S
S
Department of Chemical Pathology, University of Manchester, Hope Hospital, Salford, U.K.
White
A
A
Hoadley
M
M
Lovgren
T
T
Ratcliffe
J
J
D000324:Adrenocorticotropic Hormone / Q000097:blood*; D000911:Antibodies, Monoclonal; D003907:Dexamethasone; D005063:Europium; D005455:Fluorescent Antibody Technique; D006801:Humans; D008722:Methods; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D005063:Europium; D003907:Dexamethasone; D000324:Adrenocorticotropic Hormone