DEVELOPMENT AND VALIDATION OF A HOMOLOGOUS RADIOIMMUNOASSAY USING A BIOLOGICALLY-ACTIVE RECOMBINANT TURKEY PROLACTIN

1. A new homologous radioimmunoassay has been developed for the measur ement of turkey prolactin. 2. A 25000 kDa purified recombinant derived turkey prolactin (rtPRL), the biological activity of which was tested using a crop sac assay, was used as immunogen for the production of r abbit antiserum. In this biological test, the rtPRL was as active as t he ovinePRL. 3. The radioligand (rtPRL) was labelled with I-125 and th e assay allowed the detection of standard doses of rtPRL ranging from 400 pg/tube to 50 ng/tube. 4. No cross reaction with chicken luteinisi ng hormone and recombinant chicken growth hormone was detected. 5. The within and between assay coefficients of variability were 5.0 +/- 2.7 % and 16.3%, respectively. The overall mean recovery ratio was 1.01. 6 . The dose-response curves obtained with serial dilution of plasma and pituitary from turkey hens at different physiological stages and from male turkeys were parallel to those obtained with standard rtPRL. 7. The measured concentration of prolactin was 5 times higher in plasma f rom incubating than laying turkey hens, and the pituitaries from incub ating hens contained 2 and 4 times more prolactin than those of laying and out of lay hens or males, respectively. 8. To further assess the validity of the assay, we measured changes in plasma concentration of prolactin in turkeys following stimulation with chicken vasointestinal peptide (cVIP). A single injection of 1 or 10 mu g/kg body weight of cVIP to laying hens produced a large and rapid increase in plasma prol actin. 9. This new radioimmunoassay appears to be highly specific and sensitive for the measurement of turkey prolactin.