A method for dissociating avidin or streptavidin efficiently from a biotin derivative, thereby making it possible to isolate a target material efficiently under a mild condition in a short period, and a dissociation agent for use thereof, are provided. The method for dissociating avidin or streptavidin from a biotin derivative, includes mixing a combination of avidin or streptavidin with desthiobiotin with a water-soluble polymer to which biotin or a derivative thereof is bound.
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