Field-flow fractionation (FFF) is a family of flexible analytical fractionating techniques which have the advantage that the separation of analytes is achieved, solely through the interaction of the sample with an external, perpendicular physical field, rather than by the interaction with a stationary phase. The rapid progress in pharmaceutical biotechnology goes along with an increasing demand in potent, high-efficient analytical methods. Thus, FFF techniques are gaining increasing attention for their ability to separate and characterize populations of polymers, colloids and particles of up to about 100 microm in size. It is the intention of this review to provide an overview on common FFF techniques, to summarize inherent advantages and limitations and to introduce both established and challenging applications in the (bio)pharmaceutical field. Thereby, asymmetrical flow FFF is addressed predominantly, since it is the most versatile applicable FFF technique.