葡聚糖-卵粘蛋白亲和超滤载体的制备及应用

Preparation and Application of Dextran-Ovomucoid Affinity Ultrafiltration Carrier

为探讨葡聚糖-卵粘蛋白亲和超滤载体的制备机理及应用效果,首先用相对分子质量为200万的葡聚糖,经环氧氯丙烷活化,以卵粘蛋白为配基合成水溶性载体,然后用该载体对胰蛋白酶进行亲和超滤纯化并进行相关的紫外、红外光谱分析.结果表明:载体的活化历程为双分子亲核取代反应(SN2);合成的载体对胰蛋白酶的吸附符合Lang-muir吸附等温线方程,纯化倍数达45,活性回收率达80%;葡聚糖-卵粘蛋白载体对胰蛋白酶具有高度的亲和特异性,在亲和超滤过程中对胰蛋白酶具有很好的纯化效果.

In order to reveal the preparation mechanism of dextran-ovomucoid affinity ultrafiltration carrier and eva-luate the application effect of the carrier,first,dextran T2000（dextran with a relative molecular mass of 2×106） was employed to synthesize a water-soluble affinity carrier after the epichlorohydrin activation,with ovomucoid as the affinity ligand.Then,the changes of infrared adsorption spectrum and ultraviolet adsorption spectrum of the affinity carrier during the preparation were investigated,and pilot study on trypsin purified by the affinity ultrafiltration was performed.The results show that the activation of dextran can be described as a bimolecular nucleophilic substitution（SN2） process,that the adsorption isotherm of trypsin by dextran-ovomucoid carrier can be described by the Langmuir isotherm equation,that the purification factor and the activity recovery rate respectively reach 45 times and 80%,and that the affinity of the carrier to trypsin is of high specificity,which may result in excellent purification effect of trypsin by affinity ultrafiltration.