General strategies in the separation of proteins by high-performance liquid chromatographic methods

General fractionation strategies for the high-resolution purification of proteins are described. The impact of different separation parameters and resolution optimisation approaches with tandem-based systems on retention and recovery behaviour is reviewed. Procedures for the successful linkage of different chromatographic steps into a preferred sequence of operations are discussed in terms of the underlying principles and modus operandi of high-performance liquid chromatographic purification of proteins and related biomacromolecules.