Immobilization metal affinity chromatography(IMAC)and size-exclusive chromatography(SEC)have been widely used in the purification of recombinant protein.In order to apply the column chromatography to the separation an...Immobilization metal affinity chromatography(IMAC)and size-exclusive chromatography(SEC)have been widely used in the purification of recombinant protein.In order to apply the column chromatography to the separation and purification of the gene recombinant with histidine-tags,the column chromatographic separation characteristics of N-terminal histidine-tagged(N-AxCeSD)and C-terminal histidine-tagged(C-AxCeSD)gene recombinant protein AxCeSD,one of the subunit involved in the cellulose synthesis in Acetobacter xylinum were studied.In the ring-shaped three-dimensional structure of AxCeSD,N-terminal histidine-tags were located in the inner of ring,while C-terminal histidine-tags were located in the outer.A higher imidazole concentration was necessary for eluting the C-AxCeSD from the IMAC column due to the C-terminal histidine-tags had stronger chelating interaction with the Ni2+ on the IMAC media.Moreover,the retention time for eluting C-AxCeSD from the same SEC gel column was shorter than that for N-AxCeSD,because the larger protein homolog was formed in the C-AxCeSD solution through the inter-molecular hydrogen bonds between the C-terminal histidine-tags.展开更多
固定化金属亲和色谱(immobilized metal affinity chromatography,IMAC)在蛋白质分离纯化方面已得到广泛应用。将IMAC的固体基质部分用磁性纳米粒子(magnetic nanoparticles,MNPs)取代,可得到固定化金属亲和磁性纳米粒子(immobilized me...固定化金属亲和色谱(immobilized metal affinity chromatography,IMAC)在蛋白质分离纯化方面已得到广泛应用。将IMAC的固体基质部分用磁性纳米粒子(magnetic nanoparticles,MNPs)取代,可得到固定化金属亲和磁性纳米粒子(immobilized metal affinity magnetic nanoparticles,IMAN)。IMAN中MNPs的引入使其比IMAC具有更广泛的应用价值,近年来已广泛应用于蛋白质的分离纯化。文章重点总结了IMAN的组成、分离纯化蛋白质的原理、在蛋白质分离纯化中的应用,以及影响分离纯化蛋白质的因素,并简要评述和展望了IMAN的发展方向和应用前景。展开更多
文摘Immobilization metal affinity chromatography(IMAC)and size-exclusive chromatography(SEC)have been widely used in the purification of recombinant protein.In order to apply the column chromatography to the separation and purification of the gene recombinant with histidine-tags,the column chromatographic separation characteristics of N-terminal histidine-tagged(N-AxCeSD)and C-terminal histidine-tagged(C-AxCeSD)gene recombinant protein AxCeSD,one of the subunit involved in the cellulose synthesis in Acetobacter xylinum were studied.In the ring-shaped three-dimensional structure of AxCeSD,N-terminal histidine-tags were located in the inner of ring,while C-terminal histidine-tags were located in the outer.A higher imidazole concentration was necessary for eluting the C-AxCeSD from the IMAC column due to the C-terminal histidine-tags had stronger chelating interaction with the Ni2+ on the IMAC media.Moreover,the retention time for eluting C-AxCeSD from the same SEC gel column was shorter than that for N-AxCeSD,because the larger protein homolog was formed in the C-AxCeSD solution through the inter-molecular hydrogen bonds between the C-terminal histidine-tags.
文摘固定化金属亲和色谱(immobilized metal affinity chromatography,IMAC)在蛋白质分离纯化方面已得到广泛应用。将IMAC的固体基质部分用磁性纳米粒子(magnetic nanoparticles,MNPs)取代,可得到固定化金属亲和磁性纳米粒子(immobilized metal affinity magnetic nanoparticles,IMAN)。IMAN中MNPs的引入使其比IMAC具有更广泛的应用价值,近年来已广泛应用于蛋白质的分离纯化。文章重点总结了IMAN的组成、分离纯化蛋白质的原理、在蛋白质分离纯化中的应用,以及影响分离纯化蛋白质的因素,并简要评述和展望了IMAN的发展方向和应用前景。
