Novel plasma extraction procedure and development of a specific enzyme-immunoassay of oxytocin: application to clinical and biological investigations of small cell carcinoma of the lung.
Paraneoplastic secretion of the lactation-inducing hormone oxytocin (OT) has been reported in about 30% of cases of small cell carcinoma of the lung (SCCL). In order to investigate the role of OT in the biology of SCCL tumours, a specific enzyme-immunoassay (EIA) for OT, which can be applied to both human plasma and culture medium, has been developed. OT EIA is performed on 96-well microtiter plates coated with a rabbit polyclonal antibody (Ab) anti-OT (04). This antibody does not exhibit any significant cross-reactivity either with vasopressin (VP) or with vasotocin (VT). The immunological reaction involving Ab anti-OT is a competition between the tracer (biotinylated OT) and synthetic OT (standard curve) or OT present in biological samples. In order to limit interference induced by plasma proteins, plasma samples are filtrated by a one-step centrifugation on centricon YM-3 (cut-off 3000 Da). After plasma filtration, 90.7 +/- 5.1 (SD) % (n = 22) immunoreactive (IR) OT is recovered. The sensitivity of OT EIA is 1 pmol/L, while intra- and inter-assay coefficients of variation (CV) are around 3.41% and 2.84%, respectively. In healthy volunteers, plasma IR OT is 7.28 +/- 4.49 (SD) pmol/L (n = 32) with no gender difference. As shown by the data both from plasma of SCCL patients and from supernatants and cell contents of SCCL cell lines, this EIA procedure offers a novel, reproducible, specific and sensitive method for the measurement of IR OT.
Péqueux
C
C
University of Liège, Neuroendocrine Unit, Pathology Institute, Belgium. C.Pequeux@ulg.ac.be
Hendrick
J C
JC
Hagelstein
M T
MT
Geenen
V
V
Legros
J J
JJ
D000328:Adult; D000818:Animals; D000918:Antibody Specificity; D001667:Binding, Competitive; D019921:Biotinylation; D018288:Carcinoma, Small Cell / Q000097:blood* / Q000378:metabolism; D002498:Centrifugation; D017077:Culture Media, Conditioned; D005260:Female; D005374:Filtration; D006801:Humans; D007124:Immunoenzyme Techniques; D008175:Lung Neoplasms / Q000097:blood* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D010121:Oxytocin / Q000032:analysis* / Q000097:blood*; D011817:Rabbits; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; D010121:Oxytocin