Purification of Tetracysteine-Tagged Proteins by Affinity Chromatography Using a Non-Fluorescent, Photochemically Stable Bisarsenical Affinity Ligand

Ying, L.Q.; Branchaud, B.P., 22 (5), 987-992, Bioconjugate Chemistry, 2011

Thei article describes affinity purification of tetracysteine (CCXXCC) tagged proteins using the nonfluorescent, photochemically stable bisarsenical affinity ligand SplAsH. The Octet instrument was used to measure the binding affinity of SplAsH and tetracysteine-tagged GFP. SplAsH-biotin 5 was immobilized on Streptavidin biosensors and then incubated with purified tetracysteine-tagged GFP. Relative to the interaction between His tag and Ni-NTA, SplAsH showed >1000-fold higher affinity and thus should be much more useful for the capture and purification of low-abundance proteins.

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