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Biochemistry

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JoVE Journal Biochemistry
Removal and Replacement of Endogenous Ligands from Lipid-Bound Proteins and Allergens
 

Removal and Replacement of Endogenous Ligands from Lipid-Bound Proteins and Allergens

Article DOI: 10.3791/61780-v • 09:09 min • February 24th, 2021
February 24th, 2021 •
1, 1, 1
1Nuclear Magnetic Resonance Group, National Institute of Environmental Health Sciences

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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.

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Removal Replacement Endogenous Ligands Lipid-bound Proteins Allergens Hydrophobic Molecules Protocol Structure Immunogenicity Reverse-phase HPLC Thermal Annealing Solubilize Ligands Binding Sites Allergenicity Proteins Lipid Ligands Solubility Accessibility Loading Process Hydrophobic Ligands Insoluble Ligands Cloning Expression Purification Centrifugal Filter Unit Swing Bucket Rotor
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