Accumulation and separation of membrane-bound proteins using hydrodynamic forces
(2011) In Analytical Chemistry 83(2). p.11-604- Abstract
The separation of molecules residing in the cell membrane remains a largely unsolved problem in the fields of bioscience and biotechnology. We demonstrate how hydrodynamic forces can be used to both accumulate and separate membrane-bound proteins in their native state. A supported lipid bilayer (SLB) was formed inside a microfluidic channel with the two proteins streptavidin (SA) and cholera toxin (CT) coupled to receptors in the lipid bilayer. The anchored proteins were first driven toward the edge of the lipid bilayer by hydrodynamic forces from a flowing liquid above the SLB, resulting in the accumulation of protein molecules at the edge of the bilayer. After the concentration process, the bulk flow of liquid in the channel was... (More)
The separation of molecules residing in the cell membrane remains a largely unsolved problem in the fields of bioscience and biotechnology. We demonstrate how hydrodynamic forces can be used to both accumulate and separate membrane-bound proteins in their native state. A supported lipid bilayer (SLB) was formed inside a microfluidic channel with the two proteins streptavidin (SA) and cholera toxin (CT) coupled to receptors in the lipid bilayer. The anchored proteins were first driven toward the edge of the lipid bilayer by hydrodynamic forces from a flowing liquid above the SLB, resulting in the accumulation of protein molecules at the edge of the bilayer. After the concentration process, the bulk flow of liquid in the channel was reversed and the accumulated proteins were driven away from the edge of the bilayer. Each type of protein was found to move at a characteristic drift velocity, determined by the frictional coupling between the protein and the lipid bilayer, as well as the size and shape of the protein molecule. Despite having a similar molecular weight, SA and CT could be separated into monomolecular populations using this approach. The method also revealed heterogeneity among the CT molecules, resulting in three subpopulations with different drift velocities. This was tentatively attributed to multivalent interactions between the protein and the monosialoganglioside G(M1) receptors in the lipid bilayer.
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- author
- Jönsson, Peter LU ; Gunnarsson, Anders LU and Höök, Fredrik LU
- publishing date
- 2011-01-15
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Cholera Toxin, G(M1) Ganglioside, Hydrodynamics, Lipid Bilayers, Membrane Proteins, Microfluidic Analytical Techniques, Protein Binding, Streptavidin, Journal Article, Research Support, Non-U.S. Gov't
- in
- Analytical Chemistry
- volume
- 83
- issue
- 2
- pages
- 8 pages
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- scopus:78751491154
- pmid:21155531
- ISSN
- 1520-6882
- DOI
- 10.1021/ac102979b
- language
- English
- LU publication?
- no
- id
- 93852251-2ffd-4144-ace9-817cbded6bc6
- date added to LUP
- 2018-01-26 10:46:22
- date last changed
- 2024-04-29 03:15:08
@article{93852251-2ffd-4144-ace9-817cbded6bc6,
abstract = {{<p>The separation of molecules residing in the cell membrane remains a largely unsolved problem in the fields of bioscience and biotechnology. We demonstrate how hydrodynamic forces can be used to both accumulate and separate membrane-bound proteins in their native state. A supported lipid bilayer (SLB) was formed inside a microfluidic channel with the two proteins streptavidin (SA) and cholera toxin (CT) coupled to receptors in the lipid bilayer. The anchored proteins were first driven toward the edge of the lipid bilayer by hydrodynamic forces from a flowing liquid above the SLB, resulting in the accumulation of protein molecules at the edge of the bilayer. After the concentration process, the bulk flow of liquid in the channel was reversed and the accumulated proteins were driven away from the edge of the bilayer. Each type of protein was found to move at a characteristic drift velocity, determined by the frictional coupling between the protein and the lipid bilayer, as well as the size and shape of the protein molecule. Despite having a similar molecular weight, SA and CT could be separated into monomolecular populations using this approach. The method also revealed heterogeneity among the CT molecules, resulting in three subpopulations with different drift velocities. This was tentatively attributed to multivalent interactions between the protein and the monosialoganglioside G(M1) receptors in the lipid bilayer.</p>}},
author = {{Jönsson, Peter and Gunnarsson, Anders and Höök, Fredrik}},
issn = {{1520-6882}},
keywords = {{Cholera Toxin; G(M1) Ganglioside; Hydrodynamics; Lipid Bilayers; Membrane Proteins; Microfluidic Analytical Techniques; Protein Binding; Streptavidin; Journal Article; Research Support, Non-U.S. Gov't}},
language = {{eng}},
month = {{01}},
number = {{2}},
pages = {{11--604}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Analytical Chemistry}},
title = {{Accumulation and separation of membrane-bound proteins using hydrodynamic forces}},
url = {{http://dx.doi.org/10.1021/ac102979b}},
doi = {{10.1021/ac102979b}},
volume = {{83}},
year = {{2011}},
}