This protocol describes the removal of endogenous lipids from allergens, and their replacement with user-specified ligands through reverse-phase HPLC coupled with thermal annealing. 31P-NMR and circular dichroism allow for the rapid confirmation of ligand removal/loading, and the recovery of native allergen structure.
Foo, A. C. Y., Thompson, P. M., Mueller, G. A. Removal and Replacement of Endogenous Ligands from Lipid-Bound Proteins and Allergens. J. Vis. Exp. (168), e61780, doi:10.3791/61780 (2021).