MEP chromatography of antibody and Fc-fusion protein using aqueous arginine solution
MEP chromatography of antibody and Fc-fusion protein using aqueous arginine solution
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DOI：
10.1016/j.pep.2008.09.011
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发表时间：
2009-02-01
影响因子：
1.6
通讯作者：
Ejima, Daisuke
中科院分区：
文献类型：
--
作者：
Arakawa, Tsutomu;Kita, Yoshiko;Ejima, Daisuke
MEP HyperCel树脂，蛋白质-A模拟柱之一，被设计为在生理pH下结合抗体，并在弱酸性pH下洗脱结合的抗体。我们已经测试了精氨酸水溶液用于树脂的洗涤和洗脱。令我们惊讶的是，使用IM精氨酸溶液在pH 7.0洗脱结合的抗体和Fc融合蛋白。然后在pH 7.0下检查各种溶剂添加剂。在测试的添加剂中，尿素和精氨酸是唯一在洗脱中有效的添加剂。因此，低浓度的尿素和精氨酸有效地用于洗涤树脂。0.1-1 M的NaCl和MgCl 2以及5-20%的乙醇无效。基于这些观察结果，似乎蛋白质通过极性和疏水相互作用与MEP树脂结合，其中静电相互作用有一定贡献，其可以同时被精氨酸或尿素减少。另一方面，另一种蛋白A模拟物色谱柱Mabsorbent似乎更具非特异性和非选择性。(C)2008年爱思唯尔公司All rights reserved.
MEP HyperCel resin, one of the Protein-A mimetic columns, is designed to bind antibodies at physiological pH and elutes the bound antibodies at mildly acidic pH. We have tested aqueous arginine solution for washing and elution of the resin. To our surprise, bound antibody and Fc-fusion protein eluted at pH 7.0 using 1 M arginine solution. Various solvent additives were then examined at pH 7.0. Among the tested additives, urea and arginine were the only additives that were effective in elution. Thus, urea and arginine at low concentrations were effectively used for washing the resin. NaCl and MgCl2 at 0.1-1 M and ethanol at 5-20% were not effective. Based on these observations, it appears that protein binds to MEP resin through both polar and hydrophobic interactions with some contribution of electrostatic interaction, which can be simultaneously reduced by arginine or urea. On the other hand, Mabsorbent, another Protein-A mimetic column, appears to be more non-specific and non-selective. (C) 2008 Elsevier Inc. All rights reserved.