Monoliths
The present invention relates to a monolith for processing fluid samples, and methods of making and using the monolith. The monolith can contain certain monomers or combinations of monomers that can be polymerized to give a polymeric monolith that can efficiently self-wick fluid. The self-wicking polymeric monolith can be used as a convenient tool for point of care on site diagnostics and analytics. The monolith is easily stored and transported, comparatively cost-efficient to make, permits good detection of analyte molecules and is readily functionalizable by impregnation of and/or covalently grafting additional chemical moieties to either the whole monolith or in zones.
1. A method of fabricating a self-wicking monolith for processing a fluid sample, the method comprising:
providing at least one hydrophilic monomer and at least one linker monomer, the at least one linker monomer having two polymerizable groups spaced apart by a linker comprising at least one —C(R) 2 O— group; wherein each R is individually a hydrogen or an organic group;
obtaining a polymerizable composition by combining the at least one hydrophilic monomer and the at least one linker monomer in a porogenic solvent;
obtaining at least one additional polymerizable composition containing the at least one monomer and the at least one linker monomer in the porogenic solvent, wherein the polymerizable composition and the at least one additional polymerizable composition vary in at least one of:
the at least one hydrophilic monomer and/or the at least one linker monomer identity;
the total non-linker monomer to the at least one linker monomer ratio;
the porogenic solvent;
the concentration of the at least one hydrophilic monomer, the at least one linker monomer, and a further monomer, if present, in the solution;
the presence and identity of one or more further monomers, or
the presence and identity of an initiator;
providing the polymerizable composition and the at least one additional polymerizable composition at different locations within a mold prior to polymerization; and
polymerizing the polymerizable composition and the at least one additional polymerizable composition at the same time to form the self-wicking monolith, such that the self-wicking monolith comprises a plurality of zones, wherein different zones have different wicking properties and/or chemical properties.
2. The method of claim 1 , wherein each of the polymerizable groups of the at least one linker monomer comprises a vinylic moiety.
3. The method of claim 1 , wherein the at least one linker monomer is selected from the group consisting of:
—O—CH 2 —CH 2 —O—;
(—O—CH 2 —CH 2 —) n —O—, wherein n is selected from 2, 3, 4, or 5;
—O—CH 2 —CH(OH)—CH 2 —O—;
—O—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —O—; and
—OCH 2 —C(CH 2 O—)(CH 2 CH 3 )—CH 2 O—.
4. The method of claim 1 , wherein the at least one linker monomer is selected from the group consisting of ethyleneglycol diacrylate, ethyleneglycol dimethacrylate, tetra(ethylene glycol) dimethacrylate, tetra(ethylene glycol) diacrylate, and di(ethylene glycol) dimethacrylate.
5. The method of claim 1 , wherein the at least one hydrophilic monomer is an acrylate or methacrylate.
6. The method of claim 1 , wherein the at least one hydrophilic monomer is 2-hydroxyethyl methacrylate, 2-hydroxyacrylate, 2-hydroxyethyl acrylate, 2-hydroxypropyl methacrylate or 2-hydroxypropyl acrylate.
7. The method of claim 1 , further comprising providing at least one additional linker monomer wherein the at least one additional linker monomer is combined in the polymerizable composition.
8. The method of claim 1 , further comprising providing at least one further monomer wherein the at least one further monomer is combined in the polymerizable composition.
9. The method of claim 8 , wherein the at least one further monomer is functionalized and bears a moiety selected from the group consisting of:
a chemically reactive group suitable for reaction with a reactive group of a graftable compound to covalently graft the compound to the monolith;
a pH sensitive group;
a group suitable for direct immobilisation of an analyte;
a dye, fluorophore, chromophore, or quencher;
an immobilised protein; and
immobilised natural or artificial nucleic acid molecules.
10. The method of claim 9 , wherein the at least one further functionalized monomer comprises at least one of the following side chains or groups: amino, carboxyl, polyethylene glycol, alkyl, maleimide, succinimide, acyl halide, sulfhydryl or azide.
11. The method of claim 9 , wherein the at least one further functionalized monomer is an amino methacrylate, an amino acrylate, acrylic acid or methacrylic acid.
12. The method of claim 1 , wherein:
the porogenic solvent is able to dissolve solid monomers, or wherein the porogenic solvent is miscible with liquid monomers; and
the porogenic solvent is selected such that polymerized clusters of the at least one linker monomer precipitate from the porogenic solvent at an early point in polymerization.
13. The method of claim 1 , wherein the porogenic solvent is selected from the group consisting of:
a binary mixture containing an alkane and an alcohol;
a binary mixture containing an aromatic solvent and an alcohol;
a binary mixture containing an alcohol and a dial;
a binary mixture containing an alcohol and water;
a ternary mixture containing an alcohol, a diol and water; and
a mixture containing at least 10% (v/v) surfactant.
14. The method of claim 1 , wherein at least two zones are ordered sequentially or in parallel along an intended wicking direction of the monolith.
15. The method of claim 1 , wherein the monolith is further derivatized or modified wherein the derivatization or modification comprises at least one of the following:
a chemical hydrolysis,
a covalent grafting of a graftable compound to the monolith, or
an impregnation of the monolith with one or more components.
16. The method of claim 1 , wherein at least one of the zones is an amplification zone which is configured to facilitate amplification of a target nucleic acid sequence or whole-genome amplification in a fluid sample.
17. The method of claim 1 , wherein at least one of the zones is a clean-up zone which is configured to facilitate lysis of cells or viruses in a fluid sample.
18. The method of claim 1 , wherein at least one of the zones is a reverse transcription zone which is configured to facilitate transcription of RNA to cDNA.
19. The method of claim 1 , wherein at least one of the zones is an indication zone comprising a dye fluorophore, a chromophore, or a quencher, wherein the indication zone is configured to facilitate detection of an analyte molecule.
20. The method of claim 1 , wherein at least one of the zones is configured to retard or retain one or more components of the sample such that an analyte is separated from other components in the sample.
21. The method of claim 1 , wherein at least one of the zones is configured to facilitate a chemical transformation of one or more components of the sample.
22. The method of claim 1 , wherein the external dimensions of the monolith are:
Length: between 1 and 10 cm;
Width: between 2 and 25 mm; and
Depth: between 1 and 10 mm.
23. The method of claim 1 , wherein the total linker monomer to total hydrophilic monomer ratio for each of the polymerizable composition and the at least one additional polymerizable composition is between 1:1 and 10:1.
24. The method of claim 1 , wherein the bulk density of the monolith is between 0.15 and 0.50 g/cc.
25. The method of claim 1 , wherein the monolith comprises one or more regions having a porosity of 50 to 85%.
26. The method according to claim 1 , wherein the polymerizable composition and the at least one additional polymerizable composition abut each other in the mold; and polymerizing the abutting polymerizable composition and the at least one additional polymerizable composition occurs at the same time.
27. The method of claim 1 , wherein the plurality of polymerizable compositions at different locations within the mold are separated by dividers.
28. The method of claim 27 , further comprising removing the dividers part way through polymerization.
29. A method of fabricating a self-wicking monolith for processing a fluid sample, the method comprising:
providing at least one hydrophilic monomer and at least one linker monomer, the at least one linker monomer having two polymerizable groups spaced apart by a linker comprising at least one —C(R) 2 O— group: wherein each R is individually a hydrogen or an organic group;
obtaining a polymerizable composition by combining the at least one hydrophilic monomer and the at least one linker monomer in a porogenic solvent;
obtaining at least one additional polymerizable composition containing the at least one monomer and the at least one linker monomer in the porogenic solvent, wherein the polymerizable composition and the at least one additional polymerizable composition vary in at least one of:
the at least one hydrophilic monomer and/or the at least one linker monomer identity;
the total non-linker monomer to the at least one linker monomer ratio;
the porogenic solvent;
the concentration of the at least one hydrophilic monomer, the at least one linker monomer, and a further monomer, if present, in the solution;
the presence and identity of one or more further monomers, or
the presence and identity of an initiator;
providing the polymerizable composition and the at least one additional polymerizable composition at different locations within a mold prior to polymerization; and
polymerizing the polymerizable composition and the at least one additional polymerizable composition to form the self-wicking monolith, such that the self-wicking monolith comprises a plurality of zones, wherein different zones have different wicking properties and/or chemical properties, and wherein at least one of the zones is an amplification zone which is configured to facilitate amplification of a target nucleic acid sequence or whole-genome amplification in a fluid sample.
30. The method of claim 29 , wherein each of the polymerizable groups of the at least one linker monomer comprises a vinylic moiety.
31. The method of claim 29 , wherein the at least one linker monomer is selected from the group consisting of:
—O—CH 2 —CH 2 —O—;
(—O—CH 2 —CH 2 —) n —O—, wherein n is selected from 2, 3, 4, or 5;
—O—CH 2 —CH(OH)—CH 2 —O—;
—O—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —O—; and
—OCH 2 —C(CH 2 O—)(CH 2 CH 3 )—CH 2 O—.
32. The method of claim 29 , wherein the at least one linker monomer is selected from the group consisting of ethyleneglycol diacrylate, ethyleneglycol dimethacrylate, tetra(ethylene glycol) dimethacrylate, tetra(ethylene glycol) diacrylate, and di(ethylene glycol) dimethacrylate.
33. The method of claim 29 , wherein the at least one hydrophilic monomer is an acrylate or methacrylate.
34. The method of claim 29 , wherein the at least one hydrophilic monomer is 2-hydroxyethyl methacrylate, 2-hydroxyacrylate, 2-hydroxyethyl acrylate, 2-hydroxypropyl methacrylate or 2-hydroxypropyl acrylate.
35. The method of claim 29 , further comprising providing at least one additional linker monomer wherein the at least one additional linker monomer is combined in the polymerizable composition.
36. The method of claim 29 , further comprising providing at least one further monomer wherein the at least one further monomer is combined in the polymerizable composition.
37. The method of claim 36 , wherein the at least one further monomer is functionalized and bears a moiety selected from the group consisting of:
a chemically reactive group suitable for reaction with a reactive group of a graftable compound to covalently graft the compound to the monolith;
a pH sensitive group;
a group suitable for direct immobilisation of an analyte;
a dye, fluorophore, chromophore, or quencher;
an immobilised protein; and
immobilised natural or artificial nucleic acid molecules.
38. The method of claim 37 , wherein the at least one further functionalized monomer comprises at least one of the following side chains or groups: amino, carboxyl, polyethylene glycol, alkyl, maleimide, succinimide, acyl halide, sulfhydryl or azide.
39. The method of claim 37 , wherein the at least one further functionalized monomer is an amino methacrylate, an amino acrylate, acrylic acid or methacrylic acid.
40. The method of claim 29 , wherein:
the porogenic solvent is able to dissolve solid monomers, or wherein the porogenic solvent is miscible with liquid monomers; and
the porogenic solvent is selected such that polymerized clusters of the at least one linker monomer precipitate from the porogenic solvent at an early point in polymerization.
41. The method of claim 29 , wherein the porogenic solvent is selected from the group consisting of:
a binary mixture containing an alkane and an alcohol;
a binary mixture containing an aromatic solvent and an alcohol;
a binary mixture containing an alcohol and a diol;
a binary mixture containing an alcohol and water;
a ternary mixture containing an alcohol, a diol and water; and
a mixture containing at least 10% (v/v) surfactant.
42. The method of claim 29 , wherein at least two zones are ordered sequentially or in parallel along an intended wicking direction of the monolith.
43. The method of claim 29 , wherein the monolith is further derivatized or modified wherein the derivatization or modification comprises at least one of the following:
a chemical hydrolysis,
a covalent grafting of a graftable compound to the monolith, or
an impregnation of the monolith with one or more components.
44. The method of claim 29 , wherein at least one of the zones is a clean-up zone which is configured to facilitate lysis of cells or viruses in a fluid sample.
45. The method of claim 29 , wherein at least one of the zones is a reverse transcription zone which is configured to facilitate reverse transcription of RNA to cDNA.
46. The method of claim 29 , wherein at least one of the zones is an indication zone comprising a dye fluorophore, a chromophore, or a quencher, wherein the indication zone is configured to facilitate detection of an analyte molecule.
47. The method of claim 29 , wherein at least one of the zones is configured to retard or retain one or more components of the sample such that an analyte is separated from other components in the sample.
48. The method of claim 29 , wherein at least one of the zones is configured to facilitate a chemical transformation of one or more components of the sample.
49. The method of claim 29 , wherein the external dimensions of the monolith are:
Length: between 1 and 10 cm;
Width: between 2 and 25 mm; and
Depth: between 1 and 10 mm.
50. The method of claim 29 , wherein the total linker monomer to total hydrophilic monomer ratio for each of the polymerizable composition and the at least one additional polymerizable composition is between 1:1 to 10:1.
51. The method of claim 29 , wherein the bulk density of the monolith is between 0.15 and 0.50 Woo.
52. The method of claim 29 , wherein the monolith comprises one or more regions having a porosity of 50 to 85%.
53. The method of claim 29 , wherein the polymerizable composition and the at least one additional polymerizable composition abut each other in the mold; and polymerizing the abutting polymerizable composition and the at least one additional polymerizable composition occurs at the same time.
54. The method of claim 29 , wherein the polymerization of at least two of the compositions occurs at different times.
55. The method of claim 29 , wherein the plurality of polymerizable compositions at different locations within the mold are separated by dividers.
56. The method of claim 55 , further comprising removing the dividers part way through polymerization.
57. A method of fabricating a self-wicking monolith for processing a fluid sample, the method comprising:
providing at least one hydrophilic monomer and at least one linker monomer, the at least one linker monomer having two polymerizable groups spaced apart by a linker comprising at least one —C(R) 2 O— group; wherein each R is individually a hydrogen or an organic group;
obtaining a polymerizable composition by combining the at least one hydrophilic monomer and the at least one linker monomer in a porogenic solvent;
obtaining at least one additional polymerizable composition containing the at least one monomer and the at least one linker monomer in the porogenic solvent, wherein the polymerizable composition and the at least one additional polymerizable composition vary in at least one of:
the at least one hydrophilic monomer and/or the at least one linker monomer identity;
the total non-linker monomer to the at least one linker monomer ratio;
the porogenic solvent;
the concentration of the at least one hydrophilic monomer, the at least one linker monomer, and a further monomer, if present, in the solution;
the presence and identity of one or more further monomers, or
the presence and identity of an initiator;
providing the polymerizable composition and the at least one additional polymerizable composition at different locations within a mold prior to polymerization; and
polymerizing the polymerizable composition and the at least one additional polymerizable composition to form the self-wicking monolith, such that the self-wicking monolith comprises a plurality of zones, wherein different zones have different wicking properties and/or chemical properties, and wherein at least one of the zones is a clean-up zone which is configured to facilitate lysis of cells or viruses in a fluid sample.
58. The method of claim 57 , wherein each of the polymerizable groups of the at least one linker monomer comprises a vinylic moiety.
59. The method of claim 57 , wherein the at least one linker monomer is selected from the group consisting of:
—O—CH 2 —CH 2 —O—;
(—O—CH 2 —CH 2 —) n —O—, wherein n is selected from 2, 3, 4, or 5;
—O—CH 2 —CH(OH)—CH 2 —O—;
—O—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —O—; and
—OCH 2 —C(CH 2 O—)(CH 2 CH 3 )—CH 2 O—.
60. The method of claim 57 , wherein the at least one linker monomer is selected from the group consisting of ethyleneglycol diacrylate, ethyleneglycol dimethacrylate, tetra(ethylene glycol) dimethacrylate, tetra(ethylene glycol) diacrylate, and di(ethylene glycol) dimethacrylate.
61. The method of claim 57 , wherein the at least one hydrophilic monomer is an acrylate or methacrylate.
62. The method of claim 57 , wherein the at least one hydrophilic monomer is 2-hydroxyethyl methacrylate, 2-hydroxyacrylate, 2-hydroxyethyl acrylate, 2-hydroxypropyl methacrylate or 2-hydroxypropyl acrylate.
63. The method of claim 57 , further comprising providing at least one additional linker monomer wherein the at least one additional linker monomer is combined in the polymerizable composition.
64. The method of claim 57 , further comprising providing at least one further monomer wherein the at least one further monomer is combined in the polymerizable composition.
65. The method of claim 64 , wherein the at least one further monomer is functionalized and bears a moiety selected from the group consisting of:
a chemically reactive group suitable for reaction with a reactive group of a graftable compound to covalently graft the compound to the monolith;
a pH sensitive group;
a group suitable for direct immobilisation of an analyte;
a dye, fluorophore, chromophore, or quencher;
an immobilised protein; and
immobilised natural or artificial nucleic acid molecules.
66. The method of claim 65 , wherein the at least one further functionalized monomer comprises at least one of the following side chains or groups: amino, carboxyl, polyethylene glycol, alkyl, maleimide, succinimide, acyl halide, sulfhydryl or azide.
67. The method of claim 65 , wherein the at least one further functionalized monomer is an amino methacrylate, an amino acrylate, acrylic acid or methacrylic acid.
68. The method of claim 57 , wherein:
the porogenic solvent is able to dissolve solid monomers, or wherein the porogenic solvent is miscible with liquid monomers; and
the porogenic solvent is selected such that polymerized clusters of the at least one linker monomer precipitate from the porogenic solvent at an early point in polymerization.
69. The method of claim 57 , wherein the porogenic solvent is selected from the group consisting of:
a binary mixture containing an alkane and an alcohol;
a binary mixture containing an aromatic solvent and an alcohol;
a binary mixture containing an alcohol and a diol;
a binary mixture containing an alcohol and water;
a ternary mixture containing an alcohol, a diol and water; and
a mixture containing at least 10% (v/v) surfactant.
70. The method of claim 57 , wherein at least two zones are ordered sequentially or in parallel along an intended wicking direction of the monolith.
71. The method of claim 57 , wherein the monolith is further derivatized or modified wherein the derivatization or modification comprises at least one of the following:
a chemical hydrolysis,
a covalent grafting of a graftable compound to the monolith, or
an impregnation of the monolith with one or more components.
72. The method of claim 57 , wherein at least one of the zones is an amplification zone which is configured to facilitate amplification of a target nucleic acid sequence or whole-genome amplification in a fluid sample.
73. The method of claim 57 , wherein at least one of the zones is a reverse transcription zone which is configured to facilitate reverse transcription of RNA to cDNA.
74. The method of claim 57 , wherein at least one of the zones is an indication zone comprising a dye fluorophore, a chromophore, or a quencher, wherein the indication zone is configured to facilitate detection of an analyte molecule.
75. The method of claim 57 , wherein at least one of the zones is configured to retard or retain one or more components of the sample such that an analyte is separated from other components in the sample.
76. The method of claim 57 , wherein at least one of the zones is configured to facilitate a chemical transformation of one or more components of the sample.
77. The method of claim 57 , wherein the external dimensions of the monolith are;
Length: between 1 and 10 cm;
Width: between 2 and 25 mm; and
Depth: between 1 and 10 mm.
78. The method of claim 57 , wherein the total linker monomer to total hydrophilic monomer ratio for each of the polymerizable composition and the at least one additional polymerizable composition is between 1:1 to 10:1.
79. The method of claim 57 , wherein the bulk density of the monolith is between 0.15 and 0.50 g/cc.
80. The method of claim 57 , wherein the monolith comprises one or more regions having a porosity of 50 to 85%.
81. The method of claim 57 , wherein the polymerizable composition and the at least one additional polymerizable composition abut each other in the mold; and polymerizing the abutting polymerizable composition and the at least one additional polymerizable composition occurs at the same time.
82. The method of claim 57 , wherein the polymerization of at least two of the compositions occurs at different times.
83. The method of claim 57 , wherein the plurality of polymerizable compositions at different locations within the mold are separated by dividers.
84. The method of claim 83 , further comprising removing the dividers part way through polymerization.
85. A method of fabricating a self-wicking monolith for processing a fluid sample, the method comprising:
providing at least one hydrophilic monomer and at least one linker monomer, the at least one linker monomer having two polymerizable groups spaced apart by a linker comprising at least one —C(R) 2 O— group; wherein each R is individually a hydrogen or an organic group;
obtaining a polymerizable composition by combining the at least one hydrophilic monomer and the at least one linker monomer in a porogenic solvent;
obtaining at least one additional polymerizable composition containing the at least one monomer and the at least one linker monomer in the porogenic solvent, wherein the polymerizable composition and the at least one additional polymerizable composition vary in at least one of:
the at least one hydrophilic monomer and/or the at least one linker monomer identity;
the total non-linker monomer to the at least one linker monomer ratio;
the porogenic solvent;
the concentration of the at least one hydrophilic monomer, the at least one linker monomer, and a further monomer, if present, in the solution;
the presence and identity of one or more further monomers, or
the presence and identity of an initiator;
providing the polymerizable composition and the at least one additional polymerizable composition at different locations within a mold prior to polymerization; and
polymerizing the polymerizable composition and the at least one additional polymerizable composition to form the self-wicking monolith, such that the self-wicking monolith comprises a plurality of zones, wherein different zones have different wicking properties and/or chemical properties, and wherein at least one of the zones is a reverse transcription zone which is configured to facilitate reverse transcription of RNA to cDNA.
86. The method of claim 85 , wherein each of the polymerizable groups of the at least one linker monomer comprises a vinylic moiety.
87. The method of claim 85 , wherein the at least one linker monomer is selected from the group consisting of:
—O—CH 2 —CH 2 —O—;
(—O—CH 2 —CH 2 —) n —O—, wherein n is selected from 2, 3, 4, or 5;
—O—CH 2 —CH(OH)—CH 2 —O—;
—O—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —O—; and
—OCH 2 —C(CH 2 O—)(CH 2 CH 3 )—CH 2 O—.
88. The method of claim 85 , wherein the at least one linker monomer is selected from the group consisting of ethyleneglycol diacrylate, ethyleneglycol dimethacrylate, tetra(ethylene glycol) dimethacrylate, tetra(ethylene glycol) diacrylate, and di(ethylene glycol) dimethacrylate.
89. The method of claim 85 , wherein the at least one hydrophilic monomer is an acrylate or methacrylate.
90. The method of claim 85 , wherein the at least one hydrophilic monomer is 2-hydroxyethyl methacrylate, 2-hydroxyacrylate, 2-hydroxyethyl acrylate, 2-hydroxypropyl methacrylate or 2-hydroxypropyl acrylate.
91. The method of claim 85 , further comprising providing at least one additional linker monomer wherein the at least one additional linker monomer is combined in the polymerizable composition.
92. The method of claim 85 , further comprising providing at least one further monomer wherein the at least one further monomer is combined in the polymerizable composition.
93. The method of claim 92 , wherein the at least one further monomer is functionalized and bears a moiety selected from the group consisting of:
a chemically reactive group suitable for reaction with a reactive group of a graftable compound to covalently graft the compound to the monolith;
a pH sensitive group;
a group suitable for direct immobilisation of an analyte;
a dye, fluorophore, chromophore, or quencher;
an immobilised protein; and
immobilised natural or artificial nucleic acid molecules.
94. The method of claim 93 , wherein the at least one further functionalized monomer comprises at least one of the following side chains or groups: amino, carboxyl, polyethylene glycol, alkyl, maleimide, succinimide, acyl halide, sulfhydryl or azide.
95. The method of claim 93 , wherein the at least one further functionalized monomer is an amino methacrylate, an amino acrylate, acrylic acid or methacrylic acid.
96. The method of claim 85 , wherein:
the porogenic solvent is able to dissolve solid monomers, or wherein the porogenic solvent is miscible with liquid monomers; and
the porogenic solvent is selected such that polymerized clusters of the at least one linker monomer precipitate from the porogenic solvent at an early point in polymerization.
97. The method of claim 85 , wherein the porogenic solvent is selected from the group consisting of:
a binary mixture containing an alkane and an alcohol;
a binary mixture containing an aromatic solvent and an alcohol;
a binary mixture containing an alcohol and a did;
a binary mixture containing an alcohol and water;
a ternary mixture containing an alcohol, a diol and water; and
a mixture containing at least 10% (v/v) surfactant.
98. The method of claim 85 , wherein at least two zones are ordered sequentially or in parallel along an intended wicking direction of the monolith.
99. The method of claim 85 , wherein the monolith is further derivatized or modified wherein the derivatization or modification comprises at least one of the following:
a chemical hydrolysis,
a covalent grafting of a graftable compound to the monolith, or
an impregnation of the monolith with one or more components.
100. The method of claim 85 , wherein at least one of the zones is an amplification zone which is configured to facilitate amplification of a target nucleic acid sequence or whole-genome amplification in a fluid sample.
101. The method of claim 85 , wherein at least one of the zones is a clean-up zone which is configured to facilitate lysis of cells or viruses in a fluid sample.
102. The method of claim 85 , wherein at least one of the zones is an indication zone comprising a dye fluorophore, a chromophore, or a quencher, wherein the indication zone is configured to facilitate detection of an analyte molecule.
103. The method of claim 85 , wherein at least one of the zones is configured to retard or retain one or more components of the sample such that an analyte is separated from other components in the sample.
104. The method of claim 85 , wherein at least one of the zones is configured to facilitate a chemical transformation of one or more components of the sample.
105. The method of claim 85 , wherein the external dimensions of the monolith are:
Length: between 1 and 10 cm;
Width: between 2 and 25 mm; and
Depth: between 1 and 10 mm.
106. The method of claim 85 , wherein the total linker monomer to total hydrophilic monomer ratio for each of the polymerizable composition and the at least one additional polymerizable composition is between 1:1 to 10:1.
107. The method of claim 85 , wherein the bulk density of the monolith is between 0.15 and 0.50 g/cc.
108. The method of claim 85 , wherein the monolith comprises one or more regions having a porosity of 50 to 85%.
109. The method of claim 85 , wherein the polymerizable composition and the at least one additional polymerizable composition abut each other in the mold; and polymerizing the abutting polymerizable composition and the at least one additional polymerizable composition occurs at the same time.
110. The method of claim 85 , wherein the polymerization of at least two of the compositions occurs at different times.
111. The method of claim 85 , wherein the plurality of polymerizable compositions at different locations within the mold are separated by dividers.
112. The method of claim 111 , further comprising removing the dividers part way through polymerization.
113. A method of fabricating a self-wicking monolith for processing a fluid sample, the method comprising:
providing at least one hydrophilic monomer and at least one linker monomer, the at least one linker monomer having two polymerizable groups spaced apart by a linker comprising at least one —C(R) 2 O— group; wherein each R is individually a hydrogen or an organic group;
obtaining a polymerizable composition by combining the at least one hydrophilic monomer and the at least one linker monomer in a porogenic solvent;
obtaining at least one additional polymerizable composition containing the at least one monomer and the at least one linker monomer in the porogenic solvent, wherein the polymerizable composition and the at least one additional polymerizable composition vary in at least one of:
the at least one hydrophilic monomer and/or the at least one linker monomer identity;
the total non-linker monomer to the at least one linker monomer ratio;
the porogenic solvent;
the concentration of the at least one hydrophilic monomer, the at least one linker monomer, and a further monomer, if present, in the solution;
the presence and identity of one or more further monomers, or
the presence and identity of an initiator;
providing the polymerizable composition and the at least one additional polymerizable composition at different locations within a mold prior to polymerization; and polymerizing the polymerizable composition and the at least one additional polymerizable composition to form the self-wicking monolith, such that the self-wicking monolith comprises a plurality of zones, wherein different zones have different wicking properties and/or chemical properties, and wherein the external dimensions of the monolith are:
Length: between 1 and 10 cm;
Width: between 2 and 25 mm; and
Depth: between 1 and 10 mm.
114. The method of claim 113 , wherein each of the polymerizable groups of the at least one linker monomer comprises a vinylic moiety.
115. The method of claim 113 , wherein the at least one linker monomer is selected from the group consisting of:
—O—CH 2 —CH 2 —O—;
(—O—CH 2 —CH 2 —) n —O—, wherein n is selected from 2, 3, 4, or 5;
—OCH 2 —CH(OH)—CH 2 —O—;
—O—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —CH(OH)—CH 2 —O—; and
—OCH 2 —C(CH 2 O—)(CH 2 CH 3 )—CH 2 O—.
116. The method of claim 113 , wherein the at least one linker monomer is selected from the group consisting of ethyleneglycol diacrylate, ethyleneglycol dimethacrylate, tetra(ethylene glycol) dimethacrylate, tetra(ethylene glycol) diacrylate, and di(ethylene glycol) dimethacrylate.
117. The method of claim 113 , wherein the at least one hydrophilic monomer is an acrylate or methacrylate.
118. The method of claim 113 , wherein the at least one hydrophilic monomer is 2-hydroxyethyl methacrylate, 2-hydroxyacrylate, 2-hydroxyethyl acrylate, 2-hydroxypropyl methacrylate or 2-hydroxypropyl acrylate.
119. The method of claim 113 , further comprising providing at least one additional linker monomer wherein the at least one additional linker monomer is combined in the polymerizable composition.
120. The method of claim 113 , further comprising providing at least one further monomer wherein the at least one further monomer is combined in the polymerizable composition.
121. The method of claim 120 , wherein the at least one further monomer is functionalized and bears a moiety selected from:
a chemically reactive group suitable for reaction with a reactive group of a graftable compound to covalently graft the compound to the monolith;
a pH sensitive group;
a group suitable for direct immobilisation of an analyte;
a dye, fluorophore, chromophore, or quencher;
an immobilised protein; and
immobilised natural or artificial nucleic acid molecules.
122. The method of claim 121 , wherein the at least one further functionalized monomer comprises at least one of the following side chains or groups: amino, carboxyl, polyethylene glycol, alkyl, maleimide, succinimide, acyl halide, sulfhydryl or azide.
123. The method of claim 121 , wherein the at least one further functionalized monomer is an amino methacrylate, an amino acrylate, acrylic acid or methacrylic acid.
124. The method of claim 113 , wherein:
the porogenic solvent is able to dissolve solid monomers, or wherein the porogenic solvent is miscible with liquid monomers; and
the porogenic solvent is selected such that polymerized clusters of the at least one linker monomer precipitate from the porogenic solvent at an early point in polymerization.
125. The method of claim 113 , wherein the porogenic solvent is selected from the group consisting of:
a binary mixture containing an alkane and an alcohol;
a binary mixture containing an aromatic solvent and an alcohol;
a binary mixture containing an alcohol and a diol;
a binary mixture containing an alcohol and water;
a ternary mixture containing an alcohol, a diol and water; and
a mixture containing at least 10% (v/v) surfactant.
126. The method of claim 113 , wherein at least two zones are ordered sequentially or in parallel along an intended wicking direction of the monolith.
127. The method of claim 113 , wherein the monolith is further derivatized or modified wherein the derivatization or modification comprises at least one of the following:
a chemical hydrolysis,
a covalent grafting of a graftable compound to the monolith, or
an impregnation of the monolith with one or more components.
128. The method of claim 113 , wherein at least one of the zones is an amplification zone which is configured to facilitate amplification of a target nucleic acid sequence or whole-genome amplification in a fluid sample.
129. The method of claim 113 , wherein at least one of the zones is a clean-up zone which is configured to facilitate lysis of cells or viruses in a fluid sample.
130. The method of claim 113 , wherein at least one of the zones is a reverse transcription zone which is configured to facilitate transcription of RNA to cDNA.
131. The method of claim 113 , wherein at least one of the zones is an indication zone comprising a dye fluorophore, a chromophore, or a quencher, wherein the indication zone is configured to facilitate detection of an analyte molecule.
132. The method of claim 113 , wherein at least one of the zones is configured to retard or retain one or more components of the sample such that an analyte is separated from other components in the sample.
133. The method of claim 113 , wherein at least one of the zones is configured to facilitate a chemical transformation of one or more components of the sample.
134. The method of claim 113 , wherein the total linker monomer to total hydrophilic monomer ratio for each of the polymerizable composition and the at least one additional polymerizable composition is between 1:1 to 10:1.
135. The method of claim 113 , wherein the bulk density of the monolith is between 0.15 and 0.50 g/cc.
136. The method of claim 113 , wherein the monolith comprises one or more regions having a porosity of 50 to 85%.
137. The method of claim 113 , wherein the polymerizable composition and the at least one additional polymerizable composition abut each other in the mold; and polymerizing the abutting polymerizable composition and the at least one additional polymerizable composition occurs at the same time.
138. The method of claim 113 , wherein the polymerization of at least two of the compositions occurs at different times.
139. The method of claim 113 , wherein the plurality of polymerizable compositions at different locations within the mold are separated by dividers.
140. The method of claim 139 , further comprising removing the dividers part way through polymerization.