Since Otto Schmitt introduced the term "biomimetics" in 1957, the imitation of biological systems to develop separation methods and simulate biological processes has seen continuous growth, particularly over the past five decades. The biomimetic approach relies on the use of specific ligands-biospecific, biomimetic, or synthetic-which target biomolecules, such as proteins, antibodies, nucleic acids, enzymes, drugs, pesticides, and other bioactive analytes. This review highlights advances in biomimetic separations, focusing on biomimetic liquid chromatography (including immobilized artificial membrane chromatography, cell membrane chromatography, biomimetic affinity chromatography, weak affinity chromatography, micellar liquid chromatography, immobilized liposome chromatography, and liposome electrokinetic capillary chromatography) for the complex separation and purification of biomolecules and other important chemical compounds. It also explores their application in studying drug-receptor interactions, screening chemical permeability, absorption, distribution, toxicity, as well as predicting environmental risks. Additionally, this review discusses the application of biomimetic magnetic nanoparticles, which leverage biological membranes and proteins for drug discovery, protein purification, and diagnostics.