Isolation Purification Of Early Pregnancy Factor From Pregnant Cow Serum And Biological Assay

The early pregnancy factor (EPF) is a pregnancy protein, EPF is great significance that predicted early embryonic death and abortion, identification of embryo transfer, monitoring the status of the fetus as well as the importance of the diagnosis of esophageal cancer. EPF is an immunosuppressive protein detected in the serum in early pregnancy.we attempted to purify cow EPF from pregnant cow serum of 3-4 months. Cow EPF was purified by DEAE-Sepharose, CM-Sepharose ion-exchange chromatography, ConA-Sepharose 4B affinity chromatography and Heparin Sepharose Cl 6B affinity chromatography. Purified EPF was analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE),. EPF activity was estimated as the rosette inhibition titer by the rosette inhibition test (RIT). The result showed that purified EPF has EPF activity by the rosette inhibition test. SDS–PAGE revealed that in the final purified fraction (H-2) there were two bands of protein, and were found to be 18,300 daltons and 21,000 molecular weights, The biological activities of these proteins were stable upon heat treatment at 56℃for 30 min. Cow EPF was detected in the final purified fraction and had molecular(s) weight of 18,300 daltons and 21,000 daltons.It is prepare to further development early pregnancy factor monoclonal antibody Colloidal gold test strip of livestock abortion and infertility.