高纯度猪软骨II型胶原的制备与检测

Preparation and Analysis of Pure Type II Collagen from Porcine Articular Cartilage

探讨以猪透明软骨为原材料制备高纯度 II型胶原的方法 ,为批量化生产提供依据。以猪透明软骨为原料 ,采用盐酸胍抽提蛋白多糖 ,酸性条件下酶解 ,中间过程经多步纯化去除杂胶原、降解及变性产物 ,最后经Sepharose H.P.阴离子柱层析纯化制备出产品并以 Sigma公司产品作对照。 SDS- PAGE电泳、氨基酸成份分析和紫外最大吸收光谱的结果表明 ,所提取的 II型胶原性质符合文献报导 ,纯度比 Sigm a公司的产品高。所提取的 II型胶原为高纯度的 II型胶原 ,由于采用肉用猪作原料 ,来源丰富 ,成本低 。

This investigation was aimed at the preparation of pure type II collagen from porcine articular cartilage and the feasible method for producing type II collagen in bulk. After dispersal of the porcine articular cartilage, the proteoglycans were extracted by guanidinium hydrochloride and dissolved by pepsin in acid solvent. The contaminants including denatured and degraded protein and other collagen was removed via the repeated procedure of purification. For obtaining the purer type II collagen, the chromatography with sepharose H. P. Column was also used. The purity of the sample was compared with the type II collagen produced by Sigma Company. Both type II collagens were characterized by SDS PAGE electrophoresis, amino acid analysis and maximal violet chromatography, and all of the results accorded with the standard photograph in the references. The purity of the sample was higher than that of the product of Sigma Company. This prepared collagen of type II is a product of high purity. The raw materials are the common porcine articular cartilage, which is rich in resource and low in cost. Therefore, it is suitable to produce type II collagen in batches.