两种固定化金属螯合复合亲和膜色谱介质制备

Preparation of Two Kinds of Immobilized Metal-chelated Affinity Chromatography Membranes

以纤维素滤纸为基质 ,通过碱处理、环氧活化、偶联亚氨基二乙酸二钠、固定化Cu2 +后制得了大孔纤维素亲和膜。另外 ,在活化后的膜上通过共价交联覆盖上琼脂糖 ,制得了具有类似“三明治”结构且性能优良的复合亲和膜 ,装柱后分别制得固定化金属螯合亲和膜色谱柱。对两种亲和膜进行牛血清白蛋白等温吸附测定显示 ,两者的最大吸附量分别为 1 .1 7mg/cm2 和 1 .3 0mg/cm2 ,与传统的琼脂糖凝胶类介质吸附量相当 ,表明覆盖上的琼脂糖对提高吸附量有一定作用。依据实验结果 ,可初步推断普通纤维素滤纸可以制成性能优良的亲和膜色谱介质 ,有望在大规模分离纯化生物大分子方面得到应用。

Immobilized metal chelated affinity chromatography membranes using macroporous cellulose filter papers as stationary phase were prepared with activeagent epichorohydrin , coupling iminodiacetic acid and immobilizing Cu 2+ . Novel poly layer sandwich affinity membranes were also prepared using above filter papers with covalent cross linking agent aragose, which yielded aragose coating layer on the surface and in the porous network structure of the cellulose. After coupling iminodiacetic acid onto this aragose coated membranes, whose maximal BSA isotherm adsorption capability achieved 1.30 mg/cm 2, higher than affinity membranes which achieved 1.17 mg/cm 2, It was shown that the aragose coating layer was useful to increase adsorption capability, which is an important properties of affinity membrane, and those data obtained by affinity membranes could compare with tranditional soft gels. From these results, it could be proved that the cheap macroporous cellulose filter papers could be used to prepare affinity membranes, which are promising to use to separate and purify biomacromolecules.