Affinity Capturing and Surface Enrichment of a Membrane Protein Embedded in a Continuous Supported Lipid Bilayer

Investigations of ligand-binding kinetics to membrane proteins are hampered by their poor stability and low expression levels, which often translates into sensitivity-related limitations impaired by low signal-to-noise ratios. Inspired by affinity capturing of water-soluble proteins, which utilizes water as the mobile phase, we demonstrate affinity capturing and local enrichment of membrane proteins by using a fluid lipid bilayer as the mobile phase. Specific membrane-protein capturing and enrichment in a microfluidic channel was accomplished by immobilizing a synthesized trivalent nitrilotriacetic acid (tris-NTA)-biotin conjugate. A polymer-supported lipid bilayer containing His(6)-tagged b-secretase (BACE) was subsequently laterally moved over the capture region by using a hydrodynamic flow. Specific enrichment of His(6)-BACE in the Ni2+-NTA-modified region of the substrate resulted in a stationary three-fold increase in surface coverage, and an accompanied increase in ligand-binding response.

Simonsson Nyström, Lisa,1982Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)slisa (författare)
Burazerovic, Sabina,1976Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)bsabina (författare)
Gunnarsson, J.AstraZeneca AB (författare)
Snijder, Arjan,1971AstraZeneca AB (författare)
Geschwindner, S.AstraZeneca AB (författare)
Höök, Fredrik,1966Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)fredrikh (författare)
AstraZeneca ABChalmers tekniska högskola (creator_code:org_t)

Av författaren/redakt...: Gunnarsson, Ande ...; Simonsson Nyströ ...; Burazerovic, Sab ...; Gunnarsson, J.; Snijder, Arjan, ...; Geschwindner, S.; visa fler...; Höök, Fredrik, 1 ...; visa färre...

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