A noncompetitive enzyme immunoassay for rat prolactin.
A sensitive and specific noncompetitive rat prolactin (rPRL) enzyme immunoassay (EIA) is described. In this assay, the same rabbit anti-rPRL antibody is both adsorbed to a solid-phase support, i.e. 96-well microtiter plates and conjugated covalently to peroxidase as a tracer. PRL being sandwiched between antibody molecules, the enzymatic activity is thus proportional to the amount of rPRL concentration. This assay was found highly specific for rat PRL and displayed a sensitivity of 12.5 pg/well (0.125 ng/ml) of NIH-RP2 equivalents. The intra-assay and inter-assay coefficients of variation were less than 10% over a wide range of rPRL concentration (0.25-40 ng/ml). This rPRL-EIA permits to quantify PRL in culture media or biological samples containing up to 25% of plasma. Comparison with a radioimmunoassay revealed a good correlation (r = 0.984, the slope = 1.04). This EIA is rapid, results being obtained within 4h30 or 18h30 depending on the nature of the biological samples. The tracer, easily performed with a low cost enzyme, can be stored for very long durations. Thus, this sensitive and rapid assay provides a valuable method for measuring rPRL.
Tavernier
V
V
Synthelabo Recherche (L.E.R.S.), Département de développement chimique et pharmaceutique, Chilly Mazarin, France.
Thénot
J P
JP
Parrod
M
M
Bromet
N
N
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D007124:Immunoenzyme Techniques; D007774:Lactation; D008297:Male; D010903:Pituitary Gland, Anterior / Q000737:chemistry / Q000166:cytology; D011247:Pregnancy; D011388:Prolactin / Q000032:analysis*; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar
D011388:Prolactin