Mild conditions for releasing mono and bis-biotinylated macromolecules from immobilized streptavidin

The high affinity (kd = ∼10−15 M) of streptavidin and avidin for biotin is key to a large number of biological applications and is essentially irreversible unless the complex is exposed to harsh conditions (e.g. heat (100 °C for 10 min)), detergents, and/or denaturants which damage macromolecules. Thus, high binding affinity becomes a disadvantage when a biotinylated target must be released for further processing. This work describes relatively mild conditions that release biotin and mono- and bis-biotinylated macromolecules from immobilized streptavidin on monodispersed magnetic beads.