Chemical Engineering Department, Federal University of São Paulo (UNIFESP), Diadema, SP, 09913-030, Brazil.
School of Chemical Engineering, University of Campinas (UNICAMP), Campinas, SP, 13083-970, Brazil.
Appl Biochem Biotechnol. 2020 Jun;191(2):810-823. doi: 10.1007/s12010-019-03217-5. Epub 2019 Dec 21.
Iminodiacetic acid (IDA) is one of the chelating ligands most frequently employed in immobilized metal-ion affinity chromatography (IMAC) due to its ability to act as electron-pair donor, forming stable complexes with intermediate and borderline Lewis metal ions (electron acceptor). Thus, IDA can also be employed in ion exchange chromatography to purify positively charged proteins at neutral pH values. This study aimed to evaluate IDA as an ionogenic group (ion exchanger) immobilized on poly (ethylene vinyl alcohol) (PEVA) hollow fiber membranes for immunoglobulin G (IgG) monoclonal antibody (MAb) purification. IDA-PEVA membranes showed considerable promise for MAb purification, since IgG was recovered in eluted fractions with traces of contaminants as confirmed by Western blotting and ELISA analysis. Quantification of IgG showed that a purity of 94.2% was reached in the elution step. Breakthrough curve and batch adsorption experiments showed that the MAb dynamic binding capacity (DBC) of 3.10 mg g and the maximum adsorption capacity of 70 mg g were of the same order of magnitude as those found in the literature. The results obtained showed that the IDA-PEVA hollow fiber membrane could be a powerful adsorbent for integrating large-scale processes for purification of MAb from cell culture supernatant.
亚氨基二乙酸(IDA)是固定化金属离子亲和层析(IMAC)中最常用的螯合配体之一,因为它能够作为电子对供体,与中间和边界路易斯金属离子(电子受体)形成稳定的配合物。因此,IDA 也可以用于离子交换色谱,在中性 pH 值下纯化带正电荷的蛋白质。本研究旨在评估 IDA 作为一种离子基团(离子交换剂)固定在聚(乙烯醇)(PEVA)中空纤维膜上,用于免疫球蛋白 G(IgG)单克隆抗体(MAb)的纯化。IDA-PEVA 膜在 MAb 纯化方面显示出很大的应用前景,因为 Western blot 和 ELISA 分析证实 IgG 在洗脱部分得到回收,只有痕量的污染物。IgG 的定量表明,在洗脱步骤中达到了 94.2%的纯度。穿透曲线和批量吸附实验表明,该 MAb 的动态结合容量(DBC)为 3.10 mg/g,最大吸附容量为 70 mg/g,与文献中的值处于同一数量级。研究结果表明,IDA-PEVA 中空纤维膜可以作为一种强大的吸附剂,用于整合从细胞培养上清液中大规模纯化 MAb 的工艺。