Protocol to culture Leishmania protozoans for lipophosphoglycan extraction and purification

Lipophosphoglycan (LPG) is a macromolecule on the surface of Leishmania spp. parasites. The biochemical structure of LPG varies throughout the parasites' life cycle between proliferative (procyclic) and infective (metacyclic) stages, as well as between species and strains. Here, we outline a protocol for growing Leishmania parasites in vitro to harvest LPG. We describe steps for parasite differentiation and LPG extraction and purification. LPG has applications in medical research, such as in trained immunity and immunotherapy for cancer. For complete details on the use and execution of this protocol, please refer to dos Santos et al.¹.