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.
Claims15
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 step 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 step d).
- f) selecting from the nanoparticles obtained in step 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 step g) the population of monodisperse nanoparticles in the presence of said non aqueous solvent.
A preparation of a plasma membrane, wherein at least 60% of the proteins in said preparation are integral membrane proteins or proteins associated therewith.
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.