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Published on: December 19, 2020
Liposome Click Membrane Permeability Assay for Identifying Permeable Peptides
Tanvi J Desai1, Bahanu Habulihaz2, Joe R Cannon3
1Merck & Co., Inc., West Point, Pennsylvania, 19486, USA. tanvi.desai@merck.com.
Purpose:
To develop a novel, target agnostic liposome click membrane permeability assay (LCMPA) using liposome encapsulating copper free click reagent dibenzo cyclooctyne biotin (DBCO-Biotin) to conjugate azido modified peptides that may effectively translocate from extravesicular space into the liposome lumen.
Method:
DBCO-Biotin liposomes were prepared with egg phosphatidylcholine and cholesterol by lipid film rehydration, freeze/thaw followed by extrusion. Size of DBCO-Biotin liposomes were characterized with dynamic light scattering.
Results:
The permeable peptides representing energy independent mechanism of permeability showed higher biotinylation in LCMPA. Individual peptide permeability results from LCMPA correlated well with shifts in potency in cellular versus biochemical assays (i.e., cellular/ biochemical ratio) demonstrating quantitative correlation to intracellular barrier in intact cells.
Conclusion:
The study provides a novel membrane permeability assay that has potential to evaluate energy independent transport of diverse peptides.
