Plasma Membrane Isolation
AssigneesKatholieke Universiteit Leuven, KU LEUVEN R&D, Imec
InventorsDeepak Balaji Thimiri Govinda Raj, Liesbet Lagae, Wim Annaert, Gustaaf Borghs
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Abstract
The present invention relates to a population of monodisperse magnetic nanoparticles with a diameter between 1 and 100 nm which are coated with a layer with hydrophilic end groups. Herein the layer with hydrophilic end groups comprises an inner layer of monosaturated and/or monounsaturated fatty acids bound to said nanoparticles and bound to said fatty acids, an outer layer of a phospholipid conjugated to a monomethoxy polyethyleneglycol (PEG) comprising a hydrophilic end group, or comprises a covalently bound hydrophilic layer bound to said nanoparticles.
Claims17
A population of monodisperse positively charged magnetic nanoparticles which have a zeta potential in the range of 10-30 mV at pH 7 and a diameter between 1 and 100 nm which are coated with a layer with hydrophilic end groups, wherein said coating has an inner layer of monosaturated and/or monounsaturated fatty acids bound to said nanoparticles and bound to said fatty acids and an outer layer of a phospholipid conjugated to a monomethoxy polyethyleneglycol (PEG),
A method of preparing a monodisperse population of magnetic nanoparticles with a diameter between 1 and 100 nm which are coated with a layer with hydrophilic end groups, comprising:
- a) providing a magnetic material,
- b) applying a layer of monounsaturated and/or monosaturated fatty acid to said material by thermal decomposition under conditions to provide magnetic nanoparticles with a diameter between 1 and 100 nm,
- c) precipitating said fatty acid coated magnetic nanoparticles with an alcohol,
- d) discarding from the nanoparticles of c) the population of aggregated nanoparticles and collecting the population of monodisperse nanoparticles,
- e) applying a layer of lipids comprising a hydrophilic end group to the monodisperse nanoparticles of d),
- f) selecting from the nanoparticles obtained in e) the population of monodisperse nanoparticles in the presence of solvent,
- or instead of e) and f),
- g) replacing the fatty acid coating with a hydrophilic layer in the presence of a nonaqueous solvent (chloroform), and
- h) selecting from the nanoparticles obtained in g) the population of monodisperse nanoparticles in the presence of said non aqueous solvent.
The method of claim 7, wherein in g), said layer with hydrophilic end group is DMSA, Silane, Tetramethylammonium hydroxide (TMAOH) or ammonium chloride.
(canceled)
A preparation of a plasma membrane, wherein at least 60% of the proteins in said preparation are integral membrane proteins or proteins associated therewith.
(canceled)
The population according to claim or 2, wherein said hydrophilic end group is a phosphonate, an amine, a C1-C20 alkane, a C1-C20 alkene, a C1-C20 alkyene, azido, epoxy, NH2, COOH, unsubstituted or substituted PEG, PDP, CHO or SH.
Dependencies are inferred from the claim text.