Isolation of a bovine uterotropic placental factor and development of an enzyme immunoassay in peripheral serum

Abstract

Bovine uterotropic placental factor (bUTPF) was obtained from bovine term placentae that had been homogenized and extracted in acidic medium. The extracts were chromatographed on Sephadex G-75, a fraction named native bUTPF was obtained, and an antiserum was generated. The native bUTPF was chromatographed on Sephacryl S-300 HR, and a 230-kDa fraction was obtained. A competitive enzyme immunoassay (EIA) was developed to determine bUTPF levels during gestation. This EIA was linear between 0 and 80 μg/ml (with a detection limit of 0.2 μg/ml) and showed intraassay and interassay coefficients of variation of 11.2% and 15.5%, respectively. Bovine UTPF serum levels during gestation were determined by means of a cross-sectional study between the first month of gestation and delivery. Maximal serum levels of bUTPF were found during the first month of gestation: 7.1 ± 1.9 μg/ml (p < 0.05). In another cross-sectional study, blood samples were collected from 105 cows between 8 and 25 days postinsemination (day of insemination = Day 0). Levels of bUTPF were found to be significantly higher (p < 0.05) by Day 17 compared to Day 0 (5.8 ± 2.2 μg/ml vs. 1.2 ± 0.7 μg/ml). We conclude that bUTPF is detected by EIA in maternal peripheral serum early during gestation in the bovine species.