Flow step focusing isolates and concentrates a molecule of interest by flowing a liquid comprising a molecule of interest through a main channel having an inlet and an outlet with application of a first pressure at the inlet; applying a voltage along the channel during the flowing, wherein the voltage is configured to have a polarity such that it drives the molecule of interest in a direction opposite the flow of the liquid; controlling the first pressure and/or the voltage in a manner so as to trap and concentrate the molecule of interest in a region of the main channel; and removing the concentrated molecule of interest from the channel by recovering a portion of the liquid from a side channel diverging from the main channel, wherein the side channel is maintained at a pressure lower than the first pressure. Also disclosed is an apparatus for such.