Browsing by Author "Fogen, Dawson"

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    Engineering streptavidin and its target ligands with both infinite binding affinity and reversible binding capability
    (2015-02-06) Fogen, Dawson; Wong, Sui-Lam
    For development of reusable biosensor chips, bioreactors, protein arrays, and matrices for affinity purification of proteins, it would be ideal for streptavidin to have extremely tight binding to its target ligands (biotin and its binding tags) and at the same time to retain the feature of reversible binding capability. To achieve this objective, a streptavidin mutein (SAVSBPM32) was engineered based on a previously engineered streptavidin variant (SAVSBPM18) that can bind both biotin and its binding tag in a reversible manner. Cysteine residues were placed in strategic positions in both the streptavidin mutein and its binding tags. Disulfide bond formation allows immobilization of tagged proteins to streptavidin. Incubation with biotin in the presence of reducing agents allows stripping off tagged proteins from streptavidin.