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

BMC Veterinary Research

Jean-François Beckers¹  Claudio Canali²  Vittoria Lucia Barile³  Noelita Melo de Sousa¹  Olimpia Barbato² 

[1] Laboratory of Animal Endocrinology and Reproduction, Faculty of Veterinary Medicine, University of Liege, Liege, 4000, Belgium;Department of Biopathological Veterinary Science, Faculty of Veterinary Medicine, University of Perugia, Perugia, 06126, Italy;Animal Production Research Centre, Italian Agricultural Research Council, Monterotondo, 00016, Italy

关键词: N-terminal amino acid sequence;    Pregnancy diagnosis;    Polyclonal antiserum;    Radioimmunoassay;    Purification;    Pregnancy-associated glycoprotein;    Placenta;    Water buffalo;   

Others  :  1119549
DOI  :  10.1186/1746-6148-9-89

 received in 2012-12-18, accepted in 2013-04-22,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Barbato et al.; licensee BioMed Central Ltd.

【 图 表 】

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