Preliminary Study On Purification And Characterization Of Sheep Placental Immunoreglating Factor-Ⅰ

As the human placent and sheep placent have similarities in chemical compositions and sheep placent materials could be derived easily ,the research for sheep placent drew so many people interests at present ,but these researches only focused on the effects and clinic application of its. Sheep placent have many biologic activity factors,but we have not seen a report on which factor play an important role in the immunoreglating activity of sheep placent extraction and how to isolate one activical factor from it.To this main, we do this thesis ,the process and results of experiments as follows:Firstly,isolated and purified sheep placental immunoreglating activity factor-Ⅰfrom sheep placental extraction: After many steps including homogenization,ultracentrifugation and ultrafiltratio,sheep placental extraction with molecular weight less than 5000 Da was derived in our lab.The two components (component A and component B) were seprated from extraction by Sephadex G-50 gel filtration chromatography (column:100cm×1cm ,flow rate:0.5ml/min ). The results of which both components were inspected by high performance liquid chromatography (HPLC) showed that the component B contained the number of fators prior to that of component A, only two principal peaks. The two fractions (fraction C and fraction D) were seprated from component B by Sephadex G-25 gel filtration chromatography (column:60cm×1cm ,flow rate:0.5ml/min ). After high performance liquid chromatography (HPLC) inspection, we found that the two principal peaks of component B were well separated by this gel filtration. The two peaks were done by Sephadex G-10 gel filtration chromatography to exact salt,then were named sheep placental immunoreglating activity factor-Ⅰ(purification rate:98.89%)andⅡrespectively.As the purification rate of SPIF-Ⅰis higher than that of SPIF-Ⅱ,we made in-depth reaserach for SPIF-Ⅰ.Secondly, Spectrum and mass spectrum analysis for SPIF-Ⅰ:SPIF-Ⅰexhibited a molecular mass of 303.8 by ESI-MS, and its abstract summit of ultraviolet ray was 273nm.