凝胶过滤分离蛋白质实验条件的研究

Study on the experimental conditions in separating protein mixture by gel filtration

目的研究凝胶过滤各影响因素对蛋白质分离的影响。方法应用一般色谱方法,选择葡聚糖凝胶Sephadex G-75和几种蛋白质对装柱、洗脱液、流速和操作压等条件进行试验。结果选用不同浓度的凝胶悬液进行装柱,不同离子成分、pH的洗脱液,不同流速和操作压对几种蛋白质进行色谱分离,得到不同的结果。结论凝胶悬液较稀利于装柱,洗脱液的离子成分不直接影响分离效果,而洗脱液的pH对几种蛋白质分离产生明显影响,酸性条件下分离的效果好,适当的流速和操作压有利于提高色谱柱的分辨率,保证色谱柱的反复正常使用。

Purpose To study the effects of different elements on protein separation during gel filtration. Methods The general chromatography method, dextran Sephadex G-75 and several protein samples were used to do the assay on column packing, elution buffer, flowing rate and operational pressure. Results The different results were obtained when packed column with different gel concentrations, eluted with different ion components, pH values, flowing rate and operational pressure. Conclusion Low gel concentration works better in column packing. Ion components of elution buffer have no obvious effects on separation, while pH value of elution buffer affects the final results, good separation was got under acidic conditions, appropriate flowing rate and operational pressure improve the separation efficiency, and also they can ensure the repeated using of column.