DEVELOPMENT OF COMPOSITE OF CHITOSAN AND DEXTRAN FOR SEPARATION OF PROTEINS AND ITS APPLICATION FOR INDUSTRIAL LARGE SCALE PROTEIN SEPARATION

壳聚糖与右旋糖酐复合蛋白质分离材料的研制及其在工业化大规模蛋白质分离中的应用

基本信息

批准号：
07650926
负责人：
YOSHIDA Hiroyuki
金额：
$ 1.41万
依托单位：
Osaka Prefecture University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

Ion exchange chromatography is useful for large industrial-scale separation of proteins. A number of ion exchangers have been developed. Dextran-DEAE such as DEAE Sephadex A-50 (Pharmacia LKB Biotechnology) is a very good protein separator. However since it is very soft, it is compressed in a column even in very slow flow rate and therefore it can not be used in industrial-scale protein separation.In the present work, we have developed a composite of chitosan and dextran-DEAE to make the dextran-DEAE hard. The supporter of the dextran-DEAE was a chitosan bead, Chitopearl 2501 (Ch-2501, Fuji Spinning Co.), which was a hard gel and an adsorber of protein in itself. We let dextran penetrate into the macropores of Ch-2501 by contacting with dextran solution. Thereafter, the dextran in the macropore was cross-linked and then DEAE was introduced on the dextran network. The finished product (hereafter called Ch-Dx-DEAE-2) was hard and was not compressed in a column at all. The equilibrium iso … More therm for adsorption of BSA was affected by pH considerably. This means that Ch-Dx-DEAE-2 is feasible for separation of proteins. The isotherms were correlated well by the Langmuir equation. The saturation capacity for adsorption of BSA on Ch-Dx-DEAE-2 was 4.5,2.5, and 1.3 times larger than those of DEAE Sepharose Fast Flow (Pharmacia LKB Biotechnology), Ch-2501, and DEAE Sephadex A-50, respectively. The experimental breakthrough curve agreed well with the theoretical breakthrough curve obtained by considering the intraparticle diffusion, liquid film diffusion, and axial dispersion in the bed. The intraparticle effective diffusivities of BSA were determined by matching the experimental breakthrough curves with the theoretical ones. The diffusivity in Ch-Dx-DEAE-2 was the largest in the above ion exchangers. It was ten times larger than that in Ch-2501. BSA adsorbed on Ch-Dx-DEAE-2 could be desorbed using NaCl aqueous solution easily. The elution curve of BSA from the bed was very sharp.The value of the peak was 870 times larger than that of feed concentration of BSA in adsorption process. This is 4 times larger than that obtained using the best commercial ion exchanger which is used commonly in the world. Less

离子交换色谱法对蛋白质的大型工业规模分离很有用。已经开发了许多离子交换。 Dextran-Deae，例如Deae Sephadex A-50（Pharmacia LKB生物技术）是一种非常好的蛋白质分离剂。但是，由于它非常柔软，因此即使以非常缓慢的流量为单位，它也会在柱中压缩，因此不能在工业规模的蛋白质分离中使用。在当前的工作中，我们已经开发了壳聚糖和右旋葡萄糖的复合材料，以使葡萄糖脱氧核糖核能硬。 Dextran-Deae的支持者是壳聚糖珠2501（CH-2501，Fuji Spinning Co.），它本身就是坚硬的凝胶，并且本身是蛋白质的预言。我们通过与葡萄糖溶液接触，使葡聚糖渗透到CH-2501的大孔中。此后，大孔中的葡萄糖交联，然后在Dextran网络上引入了DEAE。最终产品（以下称为CH-DX-DEAE-2）很难，并且根本没有压缩。等效的ISO…更多用于BSA添加的热吸收受PH的影响很大。这意味着CH-DX-DEAE-2对于蛋白质的分离是可行的。等温线通过Langmuir方程很好地相关。 BSA在CH-DX-DEAE-2上添加的饱和能力分别是Deae Sepharose快速流量（Pharmacia LKB生物技术），CH-2501和DEAE SEPHADEX A-50的饱和能力。实验突破性曲线与考虑到床内的颗粒内扩散，液体膜扩散和轴向分散体获得的理论突破曲线非常吻合。通过将实验突破性曲线与理论曲线匹配，确定了BSA的内部有效差异。 CH-DX-DEAE-2中的扩散率是上述离子交换中最大的。它比CH-2501中的十倍。在CH-DX-DEAE-2上吸附的BSA可以使用NaCl水溶液来吸引。 BSA从床上的洗脱曲线非常清晰。在吸附过程中，峰值的值比BSA的进料浓度大870倍。这是使用世界上通常使用的最佳商业离子交换器获得的4倍。较少的