Optimisation of aqueous two-phase extraction of human antibodies
Optimisation of aqueous two-phase extraction of human antibodies
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DOI：
10.1016/j.jbiotec.2007.04.002
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发表时间：
2007-10-31
影响因子：
4.1
通讯作者：
Aires-Barros, M. Raquel
中科院分区：
文献类型：
--
作者：
Azevedo, Ana M.;Rosa, Paula A. J.;Aires-Barros, M. Raquel
采用表面响应法优化了聚乙二醇6000-磷酸盐双水相体系(ATPS)中人抗体的纯化条件。采用中心组合设计，考察了磷酸盐、聚乙二醇钠和氯化钠的浓度以及pH值对模拟血清培养上清液中免疫球蛋白的纯度和萃取率的影响。最大限度地将免疫球蛋白分配到上相的条件被确定为高浓度的氯化钠和聚乙二醇乙二醇酯，低浓度的磷酸盐和低pH值。用由12%聚乙二醇、10%磷酸盐、15%氯化钠组成的ATPS，在pH为6的条件下，进一步从中国仓鼠卵巢(CHO)浓缩细胞培养上清液中纯化人源单抗，富含聚乙二醇上相得率为88%，纯化倍数为4.3。该ATPS还成功地用于从杂交瘤细胞培养上清液中纯化抗体，回收率为90%，纯化倍数为4.1。(C)2007 Elsevier B.V.保留所有权利。
The purification of human antibodies in an aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) 6000 and phosphate was optimised by surface response methodology. A central composite design was used to evaluate the influence of phosphate, PEG and NaCl concentration and of the pH on the purity and extraction yield of IgG from a simulated serum medium. The conditions that maximise the partition of IgG into the upper phase were determined to be high concentrations of NaCl and PEG, low concentrations of phosphate and low pH values. An ATPS composed of 12% PEG, 10% phosphate, 15% NaCl at pH 6 was further used to purify human monoclonal antibodies from a Chinese Hamster Ovary (CHO) concentrated cell culture supernatant with a recovery yield of 88% in the upper PEG-rich phase and a purification factor of 4.3. This ATPS was also successfully used to purify antibodies from a hybridoma cell culture supernatant with a recovery yield of 90% and a purification factor of 4.1. (c) 2007 Elsevier B.V. All rights reserved.