DEVICE FOR RECOVERY AND ISOLATION OF BIOMOLECULES
The present invention relates to a device for isolating and recovering a biomolecule from a test sample. The device includes a support and at least one peelable layer deposited on at least a portion of the support. The peelable layer includes a substrate having a target component immobilized on the substrate. The device is effective for isolating and recovering a biomolecule having affinity to the target component. The present invention also relates to systems and methods of using the device. The present invention also relates to a biomolecule elution strip and related methods.
1 . A device comprising:
a support; and
at least one peelable layer deposited on at least a portion of the support, wherein said peelable layer comprises a substrate having a target component immobilized on the substrate, said target component comprising a target biomolecule and/or a target biomaterial, and
wherein said device is effective for isolating and recovering a biomolecule having affinity to the target component.
2 . The device according to claim 1 , wherein the target component is immobilized on the substrate by immobilization interactions selected from the group consisting of covalent binding, physical adsorption, and physical steric trapping.
3 . The device according to claim 1 , wherein the target component is modified to have functional groups effective to facilitate immobilization of the target component to the substrate, said functional groups being selected from the group consisting of hydrophilic groups, hydrophobic groups, carboxyl groups, amine groups, thiol groups, and the like.
4 . The device according to claim 1 , wherein the substrate comprises surface properties effective for immobilizing the target component thereto, said surface properties being characterized by the presence of functional groups selected from the group consisting of hydrophilic groups, hydrophobic groups, carboxyl groups, amine groups, gold groups, and the like.
5 . The device according to claim 1 , wherein the substrate comprises any substrate material effective to immobilize the target component thereto, said substrate material being selected from the group consisting of a polymer, a naturally occurring organic material, and the like.
6 . The device according to claim 5 , wherein the polymer is selected from the group consisting of a para-xylylene polymer (parylene), polymethyylmethacrylate (PMMA), polydimethylsiloxane (PDMS), and the like.
7 . The device according to claim 6 , wherein the para-xylylene polymer comprises Parylene A or derivatives thereof.
8 . The device according to claim 5 , wherein the naturally occurring organic material comprises nitrocellulose.
9 . The device according to claim 1 , wherein said support is configured to hold a plurality of peelable layers, wherein each of said peelable layers is independently removable from the support.
10 . The device according to claim 1 , wherein a plurality of peelable layers are deposited on the support, and wherein each peelable layer comprises either an identical or different target component.
11 . The device according to claim 1 , wherein the target biomolecule is selected from the group consisting of a protein, a peptide, a nucleic acid molecule, an aptamer, an oligonucleotide, a saccharide, a polysaccharide, a lipid, a glycolipid, a glycoprotein, a cell, and combinations thereof.
12 . The device according to claim 1 , wherein the target biomaterial is selected from the group consisting of a sol-gel, a hydrogel laden with proteins, a Matrigel, an artificially constructed scaffold with cells, and combinations thereof.
13 . The device according to claim 1 , wherein the target component has binding affinity to a biomolecule selected from the group consisting of an aptamer, a protein, a peptide, a nucleic acid molecule, an oligonucleotide, a cell-associated molecule, a saccharide, a polysaccharide, a lipid, a glycolipid, a glycoprotein, a cell, and combinations thereof.
14 . The device according to claim 1 , wherein the target component maintains its bioactivity or functionality.
15 . The device according to claim 1 , wherein the target component is selected from the group consisting of human TNF-α, human VEGF, human PDGF-BB, human PSA, human α-Thrombin, and the like.
16 . The device according to claim 1 further comprising:
a release coating deposited between the support and peelable layer, wherein said release coating is effective to facilitate separation of the peelable layer from the support.
17 . The device according to claim 16 , wherein said release coating is selected from the group consisting of a detergent/surfactant, bovine serum albumin, a thin metal layer, and the like.
18 . The device according to claim 1 , wherein the at least one peelable layer further comprises:
a secondary mechanical support layer, wherein said secondary mechanical support layer is deposited between the support and the substrate of the at least one peelable layer, said secondary mechanical support layer being effective to facilitate removal of the peelable layer from the support.
19 . The device according to claim 18 , wherein said secondary mechanical support layer comprises a para-xylylene polymer selected from the group consisting of Parylene C, Parylene AM, Parylene D, Parylene N, Parylene F, and derivatives thereof.
20 . The device according to claim 1 further comprising:
a biomolecule bound with affinity to the target component.
21 . (canceled)
22 . The device according to claim 1 , wherein the device is effective for multiplexed isolation and recovery of biomolecules from a test sample.
23 - 28 . (canceled)
29 . A method for making a device for isolating and recovering a biomolecule having affinity to a target component, said method comprising:
providing a support;
depositing a substrate onto the support, wherein the substrate is effective to immobilize a target component thereon, said target component comprising a target biomolecule and/or a target biomaterial; and
immobilizing the target component on the substrate under conditions effective to yield at least one peelable layer comprising the substrate having the target component immobilized thereon,
wherein the at least one peelable layer is removably secured to the support.
30 . The method according to claim 29 , wherein said substrate is deposited on the support by techniques selected from the group consisting of chemical vapor deposition and spin coating.
31 . The method according to claim 29 , wherein said peelable layer is removably secured to the support by electrostatic attraction and/or mechanical means.
32 . The method according to claim 29 , wherein the target component is immobilized on the substrate by immobilization interactions selected from the group consisting of covalent binding, physical adsorption, and physical steric trapping.
33 . The method according to claim 29 , wherein the target component is modified to have functional groups effective to facilitate immobilization of the target component to the substrate, said functional groups being selected from the group consisting of hydrophilic groups, hydrophobic groups, carboxyl groups, amine groups, thiol groups, and the like.
34 . The method according to claim 29 , wherein the substrate comprises surface properties effective for immobilizing the target component thereto, said surface properties being characterized by the presence of functional groups selected from the group consisting of hydrophilic groups, hydrophobic groups, carboxyl groups, amine groups, gold groups, and the like.
35 . The method according to claim 29 , wherein the substrate comprises any substrate material effective to immobilize the target component thereto, said substrate material being selected from the group consisting of a polymer, a naturally occurring organic material, and the like.
36 . The method according to claim 35 , wherein the polymer is selected from the group consisting of a para-xylylene polymer (parylene), polymethyylmethacrylate (PMMA), polydimethylsiloxane (PDMS), and the like.
37 . The method according to claim 36 , wherein the para-xylylene polymer comprises Parylene A or derivatives thereof.
38 . The method according to claim 35 , wherein the naturally occurring organic material comprises nitrocellulose.
39 . The method according to claim 29 , wherein said support is configured to hold a plurality of peelable layers, wherein each of said peelable layers is independently removable from the support.
40 . The method according to claim 29 , wherein a plurality of peelable layers are deposited on the support, and wherein each peelable layer comprises either an identical or different target component.
41 . The method according to claim 29 , wherein the target biomolecule is selected from the group consisting of a protein, a peptide, a nucleic acid molecule, an aptamer, an oligonucleotide, a saccharide, a polysaccharide, a lipid, a glycolipid, a glycoprotein, a cell, and combinations thereof.
42 . The method according to claim 29 , wherein the target biomaterial is selected from the group consisting of a sol-gel, a hydrogel laden with proteins, a Matrigel, an artificially constructed scaffold with cells, and combinations thereof.
43 . The method according to claim 29 , wherein the target component has binding affinity to a biomolecule selected from the group consisting of an aptamer, a protein, a peptide, a nucleic acid molecule, an oligonucleotide, a cell-associated molecule, a saccharide, a polysaccharide, a lipid, a glycolipid, a glycoprotein, a cell, and combinations thereof.
44 . The method according to claim 29 , wherein the target component maintains its bioactivity or functionality.
45 . The method according to claim 29 , wherein the target component is selected from the group consisting of human TNF-α, human VEGF, human PDGF-BB, human PSA, human α-Thrombin, and the like.
46 . The method according to claim 29 further comprising:
depositing a release coating between the support and peelable layer, wherein said release coating is effective to facilitate separation of the peelable layer from the support.
47 . The method according to claim 46 , wherein said release coating is selected from the group consisting of a detergent/surfactant, bovine serum albumin, a thin metal layer, and the like.
48 . The method according to claim 29 further comprising:
depositing a secondary mechanical support layer between the support and the at least one peelable layer, said secondary mechanical support layer being effective to facilitate removal of the peelable layer from the support.
49 . The method according to claim 48 , wherein said secondary mechanical support layer comprises a para-xylylene polymer selected from the group consisting of Parylene C, Parylene AM, Parylene D, Parylene N, Parylene F, and derivatives thereof.
50 . The method according to claim 29 , wherein the step of depositing at least one substrate onto at least a portion of the support further comprises:
separating the substrate into independently removable substrate strips prior to immobilizing the target component thereon,
thereby yielding a plurality of peelable layers that are independently removable from the support.
51 . The method according to claim 50 , wherein separating the substrate into independently removable substrate strips comprises using techniques selected from the group consisting of direct cutting, pre-defining perforation cutting, laser machining, photolithography patterning, and the like.
52 . The method according to claim 29 , wherein the device is in the form of a microarray comprising a plurality of peelable layers, and wherein each of the peelable layers is independently removable from the support.
53 - 59 . (canceled)