New trends in LC protein ligand screening

•A critical view of bioaffinity-based strategies for protein ligand screening.•Zonal, frontal and nonlinear chromatography with immobilized proteins is presented.•Ligand fishing with magnetic beads is an innovative tool also discussed. The diversity of small molecules available to produce truly innovative drugs associated with the wealth of known biological targets calls for key strategies in protein ligand screening. This review encompasses the recently developed bioaffinity-based strategies. A critical view of the use of zonal, frontal, and nonlinear chromatography with immobilized proteins is given. The association of these elution modes with the ligand fishing method, which uses nanomagnetics particles, is also addressed. A series of applications and how these new screening strategies can be used to determine the function, affinity, and activity parameters of proteins is discussed.