褐藻酸有限水解物作为置换剂在微异质性蛋白质置换层析中的应用

采用常规的实验仪器和通常使用的ＤＥＡＥ－纤维素５２作为层析介质,以褐藻酸有限水解物作为置换剂,对微异质性的电泳纯牛血清白蛋白进行了子交换置换层析分离。结果表明,所有７种褐藻酸水解物都能使牛血清白蛋白的洗脱曲线呈现多个洗脱峰,其中２５ｋＤａ,１５ｋＤａ,１１．５ｋＤａ和未经分离的褐藻酸水解物有更好的分离效果。

THE LIMITED HYDROLYSANTS OF ALGINATE AS A DISPLACER FOR MICRO-HETEROGENEITY PROTEIN SEPARATION IN DISPLACEMENT CHROMATOGRAPHY

The bovine serum albumin containing micro-heterogeneity was separated in ion-exchange displacement chromatography, in which the limited hydrolysants of alginate were used as displacers and the matrix was DEAE-cellulose. There were 7 kinds of the limited hydrolysants of alginate with the numeral average molecular weights of 25, 21, 15, 11.5, 7 and 5 kDa respectively and a kind of unseparated hydrolysant were used in the experiment. The results revealed that all kinds of the limited hydrolysants of alginate can separate so-called "pure" bovine serum albumin into several peaks in chromatogram. The degraded alginates with average molecular weight of 25 kDa, 15 rca, 11.5 kDa and unseparated hydrolysant can give better results. The results showed that the monomer of bovine serum albumin has about 4-6 microheterogeneity forms from comparison of chromatogram and electrophoresis scheme, besides it contains some multi-polymers and tiny unpure. The results suggested that the limited hydrolysants of alginate was a pretty displacer for protein separation in displacement chromatography.