A series of affinity media were synthesized by using the oligohistines, His Gly, (His) 3 Gly and (His) 5 Gly as the ligands and the home made monosized, non porous copolymer(CL PGMA) beads containing active epoxide gr...A series of affinity media were synthesized by using the oligohistines, His Gly, (His) 3 Gly and (His) 5 Gly as the ligands and the home made monosized, non porous copolymer(CL PGMA) beads containing active epoxide groups as the support. The affinity chromatographic behavior between IgG and the oligohistines was investigated at pH 3 0, 4 5, 7 0 and 10 0. It showed the specific interaction at pH 4 5. The dissociation constants of IgG and oligohistine media(1H, 3H, 5H) were 3 037×10 -6 , 2 459×10 -6 and 1 905×10 -6 L/mol, respectively. The dissociation constants decreased with increasing the number of histine residue in oligopeptide chains. [WT5HZ]展开更多
目的建立高效亲和色谱检测抗体融合蛋白质的方法，用于抗体及抗体融合蛋白质生产的过程控制。方法正压匀浆法装填rProtein A Sepharose，用不同pH值缓冲液分步洗脱。结果在25～1000μg／mL浓度范围内，重组人肿瘤坏死因子受体一抗体融...目的建立高效亲和色谱检测抗体融合蛋白质的方法，用于抗体及抗体融合蛋白质生产的过程控制。方法正压匀浆法装填rProtein A Sepharose，用不同pH值缓冲液分步洗脱。结果在25～1000μg／mL浓度范围内，重组人肿瘤坏死因子受体一抗体融合蛋白（TNFR—Fc）浓度和峰面积显著相关，r=0.9996；回收率范围为91％～99％，对表达TNFR—Fc的CHO工程细胞的不同培养阶段的3份上清分别进行3次重复测定，RSD在2％以内。结论该方法适用于抗体及抗体融合蛋白质生产的过程控制。展开更多
采用一步种子溶胀聚合法制备了颗粒呈单分散的交联聚甲基丙烯酸环氧丙酯树脂 ,对其环氧基的组成比进行了表征 ,并以氨基葡萄糖为配基 ,首次制备了纯化粗品伴刀豆球蛋白 A (Con A ,Ⅲ )的聚合物基质的高效亲合色谱柱。配基在树脂上的...采用一步种子溶胀聚合法制备了颗粒呈单分散的交联聚甲基丙烯酸环氧丙酯树脂 ,对其环氧基的组成比进行了表征 ,并以氨基葡萄糖为配基 ,首次制备了纯化粗品伴刀豆球蛋白 A (Con A ,Ⅲ )的聚合物基质的高效亲合色谱柱。配基在树脂上的键合量为 8.2mg g ,对纯化后的Con A 的吸附量为 13.4mg g。使用该亲合色谱介质成功地从粗品Con A中快速纯化了Con A ,电泳分析显示为一个主要的谱带 ,纯度从 15 %提高到 95 %。展开更多
文摘A series of affinity media were synthesized by using the oligohistines, His Gly, (His) 3 Gly and (His) 5 Gly as the ligands and the home made monosized, non porous copolymer(CL PGMA) beads containing active epoxide groups as the support. The affinity chromatographic behavior between IgG and the oligohistines was investigated at pH 3 0, 4 5, 7 0 and 10 0. It showed the specific interaction at pH 4 5. The dissociation constants of IgG and oligohistine media(1H, 3H, 5H) were 3 037×10 -6 , 2 459×10 -6 and 1 905×10 -6 L/mol, respectively. The dissociation constants decreased with increasing the number of histine residue in oligopeptide chains. [WT5HZ]
文摘目的建立高效亲和色谱检测抗体融合蛋白质的方法，用于抗体及抗体融合蛋白质生产的过程控制。方法正压匀浆法装填rProtein A Sepharose，用不同pH值缓冲液分步洗脱。结果在25～1000μg／mL浓度范围内，重组人肿瘤坏死因子受体一抗体融合蛋白（TNFR—Fc）浓度和峰面积显著相关，r=0.9996；回收率范围为91％～99％，对表达TNFR—Fc的CHO工程细胞的不同培养阶段的3份上清分别进行3次重复测定，RSD在2％以内。结论该方法适用于抗体及抗体融合蛋白质生产的过程控制。
文摘采用一步种子溶胀聚合法制备了颗粒呈单分散的交联聚甲基丙烯酸环氧丙酯树脂 ,对其环氧基的组成比进行了表征 ,并以氨基葡萄糖为配基 ,首次制备了纯化粗品伴刀豆球蛋白 A (Con A ,Ⅲ )的聚合物基质的高效亲合色谱柱。配基在树脂上的键合量为 8.2mg g ,对纯化后的Con A 的吸附量为 13.4mg g。使用该亲合色谱介质成功地从粗品Con A中快速纯化了Con A ,电泳分析显示为一个主要的谱带 ,纯度从 15 %提高到 95 %。
