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

DOI:
10.3791/61780-v

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09:09 min

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February 24, 2021

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, ,
1Nuclear Magnetic Resonance Group,National Institute of Environmental Health Sciences

Chapters

  • 00:04Introduciton
  • 01:13Endogenous Lipid Removal
  • 02:42Reconstitution of Apo- and Cargo-loaded Bla g 1
  • 05:07Phospholipid Cargo Removal/Loading Confirmation
  • 06:22Results: Representative Bla g 1 Purification, Lipid Removal Verification, and Recovery
  • 08:18Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Endogenous LigandsLipid-bound ProteinsAllergensReverse-phase HPLCThermal AnnealingSolubilizationBla G 1De-lipidationLyophilizationProtein Yield

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