A sensitive and specific two-site enzyme-immunoassay for human calcitonin using monoclonal antibodies.
A highly sensitive, specific and rapid two-site enzyme-immunometric assay (EIA) for the measurement of immunoreactive (ir) human calcitonin (hCT) in human plasma was developed using high-affinity monoclonal antibodies. The assay was validated in terms of sensitivity, specificity and reproducibility and its performance compared with that of a radioimmunoassay (RIA) employing a polyclonal antiserum. The sensitivity of the overnight EIA (2 pmol/l) was comparable with the long-incubation (7 days) RIA. The overnight RIA had a sensitivity of 10 pmol/l. The inter- and intra-assay variations of the EIA were less than 12%. Some related and non-related peptides were compared with synthetic hCT for cross-reactivity in the assay and were found to be negative. The mean recovery of added synthetic hCT from plasma of normal volunteers was 96%. Both RIA and EIA have been applied to the measurement of ir-hCT in normal volunteers and in patients with medullary carcinoma of the thyroid. In both groups, the level of ir-hCT measured by EIA was found to be lower than that measured by RIA, presumably due to the ability of the more specific EIA to detect only the 'mature' form of the hormone. EIA offers an attractive alternative to the more cumbersome and lengthy RIA in current usage, with the added advantage of employing a non-isotopic label.
Seth
R
R
Department of Chemical Pathology, Royal Postgraduate Medical School, London.
Motté
P
P
Kehely
A
A
Wimalawansa
S J
SJ
Self
C H
CH
Bellet
D
D
Bohuon
C
C
MacIntyre
I
I
D000469:Alkaline Phosphatase; D000911:Antibodies, Monoclonal; D002116:Calcitonin / Q000097:blood*; D002277:Carcinoma / Q000097:blood; D006801:Humans; D007124:Immunoenzyme Techniques; D011863:Radioimmunoassay; D013964:Thyroid Neoplasms / Q000097:blood; D013997:Time Factors
D000911:Antibodies, Monoclonal; D002116:Calcitonin; D000469:Alkaline Phosphatase