鹿茸硫酸软骨素蛋白聚糖的分离纯化研究

Study of isolation and identification of chondroitin sulfate proteoglycan in pliose antler

报道了用DEAE-纤维素(DE-23)离子交换柱层析从鹿茸二杠中分离、纯化及鉴定硫酸软骨素的方法。首先用适量蒸馏水浸泡鹿茸二杠并将其捣碎,离心取沉淀用盐酸胍浸提,浸提液对尿素液透析后经DEAE-纤维素(DE-23)离子交换柱层析,吸附大量的硫酸软骨素;再用含盐尿素溶液梯度洗脱、分离后,经软骨素酶消化及琼脂糖凝胶电泳,与硫酸软骨素标准品比较,证实得到的物质为纯的硫酸软骨素蛋白聚糖,其得率约为48.77%。该方法使硫酸软骨素分离纯化一步完成,大大简化了纯化步骤。

The method of purification and identification of chondroitin sulfate proteoglycan from two-branched pilose antler was reported.At first,the two-branched pilose antler was stirred in distilled water.After being centrifuged,the sediment was extracted in the carbamidine hydrochloride solution. The extract solution was exchanged by dialysis into urea solution and applied at DEAE-C(DE-23) ion exchange chromatography column,eluted with linear gradient of the salt-bearing urea solution.The purified chondroitin sulfate proteoglycan was hydrolysed by chondroitinase AC and determined by agarose gel electrophoresis.48.77 % chondroitin sulfate proteoglycan was obtained from the urea extract solution of two-branched pilose antler.This method was very quick and efficient.