Hirano Atsushi, Shiraki Kentaro, Kameda Tomoshi
Nanomaterials Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8565, Japan.
Institute of Applied Physics, University of Tsukuba, Tsukuba, Ibaraki 305-8573, Japan.
Curr Protein Pept Sci. 2019;20(1):40-48. doi: 10.2174/1389203718666171024115407.
Multimodal or mixed-mode chromatography can be used to separate various proteins, including antibodies. The separation quality and efficiency have been improved by the addition of solutes, especially arginine. This review summarizes the mechanism underlying the effects of arginine on protein elution in multimodal chromatography with neutral, anionic or cationic resin ligands; the mechanism has been investigated using experiments and molecular dynamics simulations. Arginine is effective in facilitating protein elution compared to salts and protein denaturants such as guanidine and urea. The unique elution effect of arginine can be explained by the interplay among arginine, proteins and the resin ligands. Arginine exhibits multiple binding modes for the ligands and further affinity for protein aromatic residues through its guanidinium group. These properties make arginine versatile for protein elution in multimodal chromatography. Taking into account that arginine is an aggregation suppressor for proteins but not a protein denaturant, arginine is a promising protein-eluting reagent for multimodal chromatography.
多模式或混合模式色谱法可用于分离包括抗体在内的各种蛋白质。通过添加溶质，特别是精氨酸，分离质量和效率得到了提高。本综述总结了精氨酸对使用中性、阴离子或阳离子树脂配体的多模式色谱中蛋白质洗脱影响的潜在机制；该机制已通过实验和分子动力学模拟进行了研究。与盐和蛋白质变性剂如胍和尿素相比，精氨酸在促进蛋白质洗脱方面是有效的。精氨酸独特的洗脱效果可以通过精氨酸、蛋白质和树脂配体之间的相互作用来解释。精氨酸对配体表现出多种结合模式，并通过其胍基对蛋白质芳香族残基具有进一步的亲和力。这些特性使精氨酸在多模式色谱中用于蛋白质洗脱具有通用性。考虑到精氨酸是蛋白质的聚集抑制剂而非蛋白质变性剂，精氨酸是一种很有前途的用于多模式色谱的蛋白质洗脱试剂。
