We report the induction and control of chiral recognition in liposomal membranes by the photopolymerization of diacetylenic lipids (DiynePC). The specific properties of polymerized DiynePC liposomes were characterized, and then the chiral separation performance was estimated. As the polymerization proceeds, chiral recognition to ibuprofen was induced, and its efficiency increased due to the formation of rigid nanodomains and boundary edges. Furthermore, the chiral recognition and adsorbed amount could be controlled by the ratio of rigid nanodomains, varying the composition ratio of DiynePC. Finally, the optimum condition and dominant interactions for enantioselective adsorption were clarified. Thus, our findings and results will be helpful to understand the induction of chiral recognition by polymerizable liposomes, and also become a guideline for the construction of liposomal chiral stationary phases.