Isolation And Purification Of Donkey Chorionic Gonadotropin （dCG） And Its Biological Activity

Donkey chorionic gonadotropin（dCG）,is a glycoprotein hormone found in the serum of pregnant donkeys.These hormones are produced in pregnant equine animals（horses,zebras,etc.）.The main function is to promote follicular development,at the same time,it can promote ovulation and corpus luteum formation;It has the function of promoting seminiferous tubule development and germ cell differentiation in male animals.At present,equine chorionic gonadotropin（e CG）,also known as pregnant horse serum（PMSG）,is mainly sold on the market.With the decrease of the number of horses,the supply of pregnant horse serum is insufficient,and the market gap is large.In order to study the feasibility of dCG acting on clinic,donkey serum was collected from different ages（4-7 years old）and different days of pregnancy（60,90,120 days）,and the role of ammonium sulfate salting-out combined with hydrophobic interaction chromatography in the purification of dCG was explored.The superovulation test of purified dCG and marketed e CG was carried out on 4-week-old Kunming mice to investigate the effect of dCG on mice,so as to lay a foundation for the effect of dCG on other animals.The results show that:1.The content of dCG in serum of pregnant donkey was related to the days of pregnancy.On the 90 th day of pregnancy,the average concentration of dCG activity was significantly higher than that on the 60 th and 120 th day of pregnancy.At 90 days of pregnancy,the average specific activity of 5-year-old donkeys was the highest.2.The pregnant donkey serum was roughly separated by ammonium sulfate.When the solution was 50% saturated ammonium sulfate,about 90% of the miscellaneous proteins were precipitated,and the activity loss was small,and the protein recovery rate was 88%.Then the crude separated dCG was purified by hydrophobic interaction chromatography.The purification effect is remarkable.3.In the superovulation test of mice,the effect of dCG was similar to that of e CG,which proved the possibility of dCG acting on production and life.