Dept. of Chemical and Biological Engineering, Missouri University of Science and Technology, Rolla, MO 65409, USA.
Biotechnol Prog. 2010 Jan-Feb;26(1):159-67. doi: 10.1002/btpr.287.
A recombinant human antibody expressed in corn was purified using aqueous two-phase extraction. The antibody was an immunoglobulin G fully unglycosylated. Using systems of different compositions and/or pHs in each of one or two partitioning stages followed by one more stage in which the antibody was precipitated at the liquid/liquid interface facilitated the removal of different impurities in each stage. The best system yields a product 72% pure (22-fold purification) with a yield of 49%. The optimum extraction was done in two partitioning stages followed by an interfacial precipitation stage using poly(ethylene)glycol/potassium phosphate systems. NaCl was added to the first stage to eliminate large molecular weight impurities. The pH in the first stage was kept at 6 but a pH of 8 was used in the second stage and in the precipitation stage.
在玉米中表达的重组人抗体使用双水相萃取进行纯化。该抗体是完全未糖基化的免疫球蛋白 G。在一个或两个分配阶段的每一个阶段使用不同组成和/或 pH 的系统，然后在另一个阶段在液/液界面处沉淀抗体，有助于在每个阶段去除不同的杂质。最佳系统的产率为 49%，产物纯度为 72%（纯化 22 倍）。使用聚乙二醇/磷酸钾系统在两个分配阶段后进行界面沉淀阶段进行最佳萃取。在第一阶段添加 NaCl 以去除高分子量杂质。第一阶段的 pH 值保持在 6，但第二阶段和沉淀阶段的 pH 值为 8。
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