| Literature DB >> 19362151 |
Mutsumi Futatsumori-Sugai1, Ryota Abe, Masato Watanabe, Motonori Kudou, Tadashi Yamamoto, Daisuke Ejima, Tsutomu Arakawa, Kouhei Tsumoto.
Abstract
FLAG-tag is one of the commonly used purification technologies for recombinant proteins. An antibody, M2, specifically binds to the FLAG-tag whether it is attached to N- or C-terminus of proteins to be purified. The bound proteins are generally eluted by competition with a large excess of free FLAG peptide. This requires synthetic FLAG peptide and also removal of bound FLAG peptide for M2 column regeneration. We have shown before that arginine at mild pH can effectively dissociate protein-protein or protein-ligand interactions, e.g. in Protein-A, antigen and dye-affinity chromatography. We have tested here elution of FLAG-fused proteins by arginine for columns of M2-immobilized resin using several proteins in comparison with competitive elution by FLAG peptide or low pH glycine buffer. Active and folded proteins were successfully and effectively eluted using 0.5-1M arginine at pH 3.5-4.4, as reported in this paper.Entities:
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Year: 2009 PMID： 19362151 DOI： 10.1016/j.pep.2009.03.012
Source DB: PubMed Journal: Protein Expr Purif ISSN： 1046-5928 Impact factor: 1.650