Purification of pregnancy-associated glycoproteins from late-pregnancy Bubalus bubalis placentas and development of a radioimmunoassay for pregnancy diagnosis in water buffalo females

BMC Vet Res. 2013 May 1:9:89. doi: 10.1186/1746-6148-9-89.

Abstract

Background: Pregnancy-associated glycoproteins (PAGs) were first described as placental antigens present in the blood serum of the mother soon after implantation. Here, we describe the purification of several pregnancy-associated glycoproteins from water buffalo placenta (wbPAGs). A specific radioimmunoassay (RIA) was developed for early pregnancy diagnosis in buffalo species.

Results: Amino-terminal microsequencing of immunoreactive placental proteins allowed the identification of eleven wbPAGs sequences [Swiss-Prot accession numbers: P86369 to P86379]. Three polyclonal antisera (AS#858, AS#859 and AS#860) were raised in rabbits against distinct wbPAG fractions. A new RIA (RIA-860) was developed and used to distinguish between pregnant (n=33) and non-pregnant (n=26) water buffalo females.

Conclusions: Our results confirmed the multiplicity of PAG expression in buffalo placenta. In addition, the RIA-860 system was shown to be sensitive, linear, reproducible, accurate and specific in measuring PAG concentrations in buffalo plasma samples from Day 37 of gestation onwards.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Buffaloes / metabolism*
  • Female
  • Glycoproteins / blood
  • Glycoproteins / immunology
  • Glycoproteins / isolation & purification*
  • Immune Sera / immunology
  • Molecular Sequence Data
  • Placenta / chemistry*
  • Pregnancy
  • Pregnancy Proteins / blood
  • Pregnancy Proteins / immunology
  • Pregnancy Proteins / isolation & purification*
  • Pregnancy Tests, Immunologic / methods
  • Pregnancy Tests, Immunologic / veterinary*
  • Rabbits / immunology
  • Radioimmunoassay / methods
  • Radioimmunoassay / veterinary*
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Glycoproteins
  • Immune Sera
  • Pregnancy Proteins

Associated data

  • SWISSPROT/P86369
  • SWISSPROT/P86370
  • SWISSPROT/P86371
  • SWISSPROT/P86372
  • SWISSPROT/P86373
  • SWISSPROT/P86374
  • SWISSPROT/P86375
  • SWISSPROT/P86376
  • SWISSPROT/P86377
  • SWISSPROT/P86378
  • SWISSPROT/P86379