ISOLATION AND PURIFICATION OF PREGNANCY-ASSOCIATED GLYCOPROTEINS

The aim of these studies was to develop a protocol for the isolation and purification pregnancy-associated glycoproteins (PAG) from cotyledons placenta pregnant cows. As result this work, processes parameters PAG were determined, including: extraction in buffer system, two-stage precipitation with ammonium sulfate (40% 80% saturation) target product using ion-exchange chromatography Q Sepharose Fast Flow. resulting having molecular weight 55 kD identified by MALDI-TOF mass-spectrometry analysis as "Pregnancy-associated glycoprotein 1". immunoblotting, presence proteins kDa obtained preparation, reacting previously monoclonal antibodies (MABs) specific PAG1 confirmed. To determine antigenic properties purified PAG, laboratory animals immunized. It found that immunization efficiently elicits antibodies. These antibodies, after proper procedure, can be used components immunological test systems determining pregnancy