column chromatography separation of crude and purified protein in cobratoxin and analysis of anti-tumor activity of its fractions

Objective:To separate crude protein and purified protein in cobratoxin and analyze anti-tumor activity of its fractions.Methods:The crude protein and purified protein in cobratoxin was separated by ion exchange chromatography.Qualitative study was made on each fraction by means of analysis by reverse-phase chromatography,content determination and anti-tumor activity test.Results: Four elution peaks were obtained when a linear gradient elution was applied to crude protein and purified protein in cobratoxin by adding sodium chloride(0-0.5 mol·L-1) to Tris-HCl buffer(20 mmol·L-1,pH 8.0) on DEAE-sephacel ion-exchange column.Among the four elution peaks,the first one showed the highest anti-tumor activity.It contained two types of proteins,which differed in hydrophobicity and ultraviolet absorption,while the other three elution peaks contained single protein respectively.Conclusion: The fractions from cobratoxin have some inhibitory effects on tumor cell lines cultured in vitro.