Abstract 2896 Switchable affinity streptavidin muteins for purification and proteomics applications

Due to the high affinity and high specificity of its interaction with biotin, streptavidin is widely used in basic research and biotechnological applications to capture biotinylated molecules from complex mixtures. However, this high affinity also means that the interaction is essentially irreversible under non-denaturing conditions over practical timescales, precluding its use when recovery of the intact target molecule or reusability of the matrix is required. To overcome this limitation, we have engineered a series of streptavidin muteins in which the reduction of a non-natural disulfide bond can increase the rates of dissociation by up to four orders of magnitude.