Purification And Physicochemical Properties Of Low Molecular Weight Hepatocyte Growth Factor Is Discussed

PARTⅠ PURIFICATION OF LOW MOLECULAR WEIGHT HEPATOPOEITIN ABSTRACT[Objective] Purification of low molecular weight hepatopoeitin[Methods] 1. With Shim-pack Diol 150 column, hepatopoeitin is separated by high performance liquid chromatography(HPLC). After the separation, the active fractions of low molecular weight hepatopoeitin are detected by PI(proliferation index)with the methods of stimulating DNA synthesis in HepG2 cell and QGY cell.2. Inactive components of unpurified solution are removed by continuous precipitations in - 20 ℃ acetone.3. Removing inactive components, we obtain pure active fractions of low molecular weight hepatopoeitin with Shim-pack Diol 150 again, then evaluate molecular weight in Tricine-SDS-PAGE.[Results] 1. Hepatopoeitin is separated to nine fractions by HPLC.The first 1 to 4 fractions show positive activity,and the activity of the fourth fraction is the highest. 2.The main fraction of precipitation in 1:12(V/V) acetone precipitations is the fourth fraction in HPLC which shows the highest activity.3.The final purified low molecular weight hepatopoeitin is obtained by Shim-pack Diol 150 column with 1:12(v/v) acetone precipitations. It shows a zone about 6,540 dalton in Tricine-SDS-PAGE.