一种通过对溶解于盐酸胍中的化合物进行乙醇沉淀，从生物样品中分离肝素种类的方法。

A method for separation of heparin species from biological samples by ethanol precipitation of compounds solubilized in guanidine hydrochloride.

作者信息

Ruggiero M, Caldini R, Chevanne M, Melli M, Pacini S, Gulisano M, Vannucchi S

机构信息

Department of Experimental Pathology and Oncology, University of Firenze, Italy.

出版信息

Ital J Anat Embryol. 2001 Jan-Mar;106(1):35-46.

PMID:11410997

Abstract

In this paper we describe a procedure to determine glycosaminoglycan and oligosaccharide composition of biological samples such as cell cultures or tissue explants. We demonstrate that heparin species of different molecular mass can be easily fractionated by sequential ethanol precipitation in 4.0 M guanidine hydrochloride. We studied by gradient polyacrylamide gel electrophoresis fractionation of standard heparin and heparin-derived oligosaccharides by anion-exchange chromatography on DEAE-Sephacel resin eluted by increasing concentration of guanidine hydrochloride. The use of guanidine salts followed by sequential precipitation by increasing ethanol concentration allowed recovery of heparin and heparin-derived oligosaccharides.

摘要

在本文中，我们描述了一种用于确定生物样品（如细胞培养物或组织外植体）中糖胺聚糖和寡糖组成的方法。我们证明，不同分子量的肝素种类可以通过在4.0 M盐酸胍中依次进行乙醇沉淀而轻松分离。我们通过梯度聚丙烯酰胺凝胶电泳研究了标准肝素和肝素衍生的寡糖在DEAE-葡聚糖凝胶树脂上进行阴离子交换色谱分离，用逐渐增加浓度的盐酸胍洗脱。使用胍盐，随后通过增加乙醇浓度依次沉淀，可以回收肝素和肝素衍生的寡糖。