1-22. (canceled)
23. A method of separating a target molecule by solid phase adsorption comprising passing a sample containing said target molecule through a chromatography device loaded with a solid phase matrix comprising dense mineral oxide solid supports comprising:
(a) a mineral oxide matrix having an external surface and pores, wherein the pores have a pore volume which is less than 30% of the total volume of the mineral oxide matrix, and
(b) an interactive polymer network which fills the pores and is coated on the surface of the mineral oxide matrix, so that subsequent interaction with macromolecules occurs on the external surface area of the support.
24. The method of claim 23, wherein the target molecule is a biological molecule.
25. A method for separating a desired biological molecule from a sample solution containing the same comprising the steps of:
a) loading a chromatography device with a chromatography bed comprised of dense mineral oxide solid supports comprising
i) a mineral oxide matrix having a pore volume which is less than 30% of the total volume of the mineral oxide matrix, and
ii) an interactive polymer network which is rooted in pores and on the surface of the mineral oxide matrix, wherein the interactive polymer network is functionalized to have affinity for the desired biological molecule;
b) feeding the sample solution containing said desired biological molecule into the chromatography device, whereby the desired biological molecule is adsorbed to the dense mineral oxide solid supports;
c) washing the chromatography device with a washing buffer and discharging undesired components and impurities of the sample solution from the chromatography device;
d) feeding an eluting buffer into the chromatography device, wherein said eluting buffer causes the desired biological molecule to be released from the dense mineral oxide solid supports; and
e) collecting the desired biological molecule.
26. The method of claim 25, wherein the dense mineral oxide solid supports have a density in the range of about 2.1 to about 11.
27. The method of claim 25, wherein said dense mineral oxide solid supports have a particle size in the range of about 5 \u03bcm to about 500 \u03bcm.
28. The method of claim 27, wherein the particle size is about 10 \u03bcm to about 100 \u03bcm.
29. The method of claim 25, wherein the mineral oxide matrix is comprised of titania, zirconia, yttria, ceria, hafnia, tantalia, or mixtures thereof.
30. The method of claim 28, wherein the mineral oxide matrix is comprised of titania, zirconia, yttria, ceria, hafnia, tantalia, or mixtures thereof.
31. The method of claim 25, wherein the interactive polymer network comprises a soluble organic polymer or a mixture of soluble organic polymers crosslinked in place with the mineral oxide matrix.
32. The method of claim 25, wherein the interactive polymer network comprises monomers, bifunctional monomers, or mixtures thereof copolymerized in place with the mineral oxide matrix.
33. The method of claim 25, wherein the desired biological molecule is a macromolecule.
34. The method of claim 33, wherein the macromolecule is a polysaccharide, a plasmid, a nucleic acid, a polynucleotide, or a protein aggregate.
35. The method of claim 25, wherein the desired biological molecule is a bioparticle.
36. The method of claim 35, wherein the bioparticle is a virus, a viral vector, a membrane protein, or a cellular structure.
37. The method of claim 25, wherein the chromatography device is a packed bed column, a fluidized bed column, or a continuous stirred tank.
38. A fluidized bed chromatography method for separating a desired biological molecule from a sample solution containing the same comprising the steps of:
a) loading a fluidized bed column with a chromatography bed comprised of dense mineral oxide solid supports comprising
i) a mineral oxide matrix having a pore volume which is less than 30% of the total volume of the mineral oxide matrix, and
ii) an interactive polymer network which is rooted in pores and on the surface of the mineral oxide matrix, wherein the interactive polymer network is functionalized to have affinity for the desired biological molecule;
b) feeding an initial buffer into said fluidized bed column at a linear velocity which causes the dense mineral oxide solid supports to form a fluidized bed;
c) feeding the sample solution containing said desired biological molecule into the fluidized bed column at a linear velocity which maintains the dense mineral oxide solid supports in the fluidized bed, whereby the desired biological molecule is adsorbed to the dense mineral oxide solid supports;
d) washing the chromatography device with a washing buffer and discharging undesired components and impurities of the sample solution from the fluidized bed column device;
e) feeding an elution buffer into the fluidized bed column, wherein said elution buffer causes the desired biological molecule to be released from the dense mineral oxide solid supports; and
f) collecting the desired biological molecule eluted from the fluidized bed column.
39. The method of claim 38, wherein the dense mineral oxide solid supports have a density in the range of about 2.1 to about 11.
40. The method of claim 38, wherein said dense mineral oxide solid supports have a particle size in the range of about 5 \u03bcm to about 500 \u03bcm.
41. The method of claim 40, wherein the particle size is about 10 \u03bcm to about 100 \u03bcm.
42. The method of claim 38, wherein the mineral oxide matrix is comprised of titania, zirconia, yttria, ceria, hafnia, tantalia, or mixtures thereof.
43. The method of claim 41, wherein the mineral oxide matrix is comprised of titania, zirconia, yttria, ceria, hafnia, tantalia, or mixtures thereof
44. The method of claim 38, wherein the interactive polymer network comprises a soluble organic polymer or a mixture of soluble organic polymers crosslinked in place with the mineral oxide matrix.
45. The method of claim 38, wherein the interactive polymer network comprises monomers, bifunctional monomers, or mixtures thereof copolymerized in place with the mineral oxide matrix.
46. The method of claim 38, wherein the desired biological molecule is a macromolecule.
47. The method of claim 46, wherein the macromolecule is a polysaccharide, a plasmid, a nucleic acid, a polynucleotide, or a protein aggregate.
48. The method of claim 38, wherein the desired biological molecule is a bioparticle.
49. The method of claim 48, wherein the bioparticle is a virus, a viral vector, a membrane protein, or a cellular structure.
50. The fluidized bed chromatography method of claim 38, wherein the linear velocity is within the range of 100 cmhour to 3000 cmhour.
51. A method for preparing dense mineral oxide solid supports which comprises:
(a) preparing a mixture of particles of at least one mineral oxide;
(b) forming a mineral oxide matrix from said mixture;
(c) sintering the resulting mineral oxide matrix at a high temperature which melts subparticles in the mineral oxide matrix, wherein the sintering reduces the pore volume of the mineral oxide matrix to less than 30% of the total volume of the mineral oxide matrix; and
(d) forming an interactive polymer network rooted in the pores and on the surface of the resulting sintered mineral oxide matrix.
52. The method of claim 51, wherein the mineral oxide is selected from the group consisting of titania, zirconia, yttria, ceria, hafnia, tantalia, or mixtures thereof.
53. The method of claim 51, wherein the particles of mineral oxide have a particle size of in the range of 0.1 \u03bcm to 15 \u03bcm.
54. The method of claim 53, wherein the particles of mineral oxide have a particle size of 0.1 \u03bcm to 3 \u03bcm.
55. The method of claim 51, wherein the dense mineral oxide solid supports have a rough surface, and wherein the particles of mineral oxide have a particle size of 3 \u03bcm to 15 \u03bcm.
56. The method of claim 51, wherein the beads are formed by a sol-gel process, a spray drying process, or an emulsion-polycondensation process.
57. The method of claim 51, wherein the interactive polymer network is comprised of monomers, bifunctional monomers, or mixtures thereof copolymerized in place with the mineral oxide matrix.
58. The method of claim 51, wherein the interactive polymer network is comprised of a soluble organic polymer or a mixture of soluble organic polymers crosslinked in place with the mineral oxide matrix.
59-63. (canceled)
64. The method of claim 23, wherein the pore volume is 5% to 25% of the total volume of the mineral oxide matrix.
65. The method of claim 23, wherein the pore volume is 5% to 15% of the total volume of the mineral oxide matrix.
66. The method of claim 25, wherein the pore volume is 5% to 25% of the total volume of the mineral oxide matrix.
67. The method of claim 25, wherein the pore volume is 5% to 15% of the total volume of the mineral oxide matrix.
68. The method of claim 38, wherein the pore volume is 5% to 25% of the total volume of the mineral oxide matrix.
69. The method of claim 38, wherein the pore volume is 5% to 15% of the total volume of the mineral oxide matrix.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A metal substrate coated at least partially with a layered structure, said layered structure comprising:
an intermediate layer deposited on said substrate and a tetrahedral carbon layer deposited on said intermediate layer,
wherein said intermediate layer comprises at least one amorphous carbon layer having a Young’s modulus lower than 200 GPa, and
wherein said tetrahedral carbon layer is a non-hydrogenated tetrahedral carbon layer having a Young’s modulus higher than 200 GPa.
2. A substrate according to claim 1, wherein said layered structure comprises a number of periods, wherein each period comprises an intermediate layer comprising at least one amorphous carbon layer having a Young’s modulus lower than 200 GPa and a non-hydrogenated tetrahedral carbon layer having a Young’s modulus higher than 200 GPa, and wherein said number of periods is between 2 and 100.
3. A substrate according to claim 1, wherein said tetrahedral carbon layer has a Young’s modulus ranging between 200 and 800 GPa.
4. A substrate according to claim 1, wherein said tetrahedral carbon layer has a hardness higher than 20 GPa.
5. A substrate according to claim 1, wherein said tetrahedral carbon layer has a fraction of sp3 bonded carbon higher than 30%.
6. A substrate according to claim 1, wherein said tetrahedral carbon layer has a fraction of sp3 bonded carbon higher than 80%.
7. A substrate according to claim 1, wherein said tetrahedral carbon layer is doped with a metal.
8. A substrate according to claim 1, wherein said amorphous carbon layer is selected from the group consisting of amorphous hydrogenated carbon (a-C:H) provided with Si and O and amorphous hydrogenated carbon (a-C:H).
9. A substrate according to claim 8, wherein said amorphous carbon layer provided with Si and O comprises a first interpenetrating network of predominantly sp3 bonded carbon in a diamond-like carbon network stabilized by hydrogen, and a second interpenetrating network of silicon stabilized by oxygen.
10. A substrate according to claim 1, wherein said amorphous carbon layer is doped with at least one metal.
11. A substrate according to claim 1, wherein said layered structure further comprises an adhesion promoting layer deposited on said substrate before deposition of said intermediate layer.
12. A substrate according to claim 11, wherein said adhesion promoting layer comprises at least one layer, wherein said at least one layer comprises at least one element of the group consisting of silicon, an element from group IVB of the periodic table, an element from group VB of the periodic table, and an element from group VIB of the periodic table.
13. A substrate according to claim 11, wherein said adhesion promoting layer comprises at least one metal layer, said at least one metal layer comprising at least one element of the group consisting of silicon, an element from group IVB of the periodic table, an element from group VB of the periodic table, and an element from group VIB of the periodic table.
14. A substrate according to claim 11, wherein said adhesion promoting layer comprises at least one layer selected from form the group consisting of carbides, nitrides, carbonitrides, oxycarbides, oxynitrides, oxycarbonitrides of at least one element of the group consisting of silicon, an element from group IVB of the periodic table, an element from group VB of the periodic table, and an element from group VIB of the periodic table.
15. A substrate according to claim 11, wherein said adhesion promoting layer comprises a combination of at least one metal layer of a metal selected from the group consisting of silicon, an element from group IVB of the periodic table, an element from group VB of the periodic table, and an element from group VIB of the periodic table and at least one layer of a carbide, a nitride, a carbonitride, an oxycarbide, an oxynitride, an oxycarbonitride of a metal selected from the group consisting of silicon, an element from group IVB of the periodic table, an element from group VB of the periodic table, and an element from group VIB of the periodic table.
16. A substrate according to claim 1, wherein said layered structure further comprises a top layer, and wherein said top layer is deposited on said tetrahedral carbon layer.
17. A substrate according to claim 16, wherein said top layer is selected from the group consisting of amorphous hydrogenated carbon (a-C:H); amorphous hydrogenated carbon (a-C:H) doped with one or more of elements O, N andor F; amorphous hydrogenated carbon (a-C:H) provided with Si and O and being metal doped or doped with one or more of the elements O, N andor F; and metal doped hydrogenated carbon.