Detection and assay devices and methods of making and using the same
An article such as a biomolecular detector or biosensor having a nonfouling surface thereon includes: (a) a substrate having a surface portion; (b) a linking layer on the surface portion; and (c) a polymer layer formed on the linking layer; and (d) a first member of a specific binding pair (e.g., a protein, peptide, antibody, nucleic acid, etc.) bound to the polymer layer. Methods of making and using the articles are also described.
1. A method of non-covalently binding a first member of a specific binding pair to a non-fouling polymer layer, comprising:
(a) providing the non-fouling polymer layer, which is at least macroscopically dry and retains water of hydration, wherein said non-fouling polymer layer comprises brush molecules, each brush molecule comprising:
(i) a polymeric stem comprising monomer core groups having at least one protein-resistant head group coupled thereto, wherein said at least one protein-resistant head group comprises oligo(ethylene glycol)methyl methacrylate (OEGMA), and
(ii) a plurality of branches comprising said at least one protein-resistant head group projecting from said stem;
(b) depositing the first member of the specific binding pair on the non-fouling polymer layer without activation or deactivation of the brush molecules, wherein the first member of a specific binding pair is directly, non-covalently bound to the non-fouling polymer layer; and
(c) drying the non-fouling polymer layer containing the non-covalently bound first member of a specific binding pair for a time sufficient for the non-fouling polymer layer to be at least macroscopically dry,
and wherein a second member of the specific binding pair is a constituent of a biological fluid.
2. The method of claim 1 , wherein the depositing comprises non-contact printing.
3. The method of claim 1 , wherein the depositing comprises direct contact quill printing.
4. The method of claim 1 , wherein the depositing comprises dip pen lithography.
5. The method of claim 1 , wherein the depositing comprises piezoelectric printing.
6. The method of claim 1 , wherein the depositing comprises a technique selected from the group consisting of microprinting, microstamping, and any combination thereof.
7. The method of claim 1 , wherein the brush molecules are from 2-200 nanometers (nm) in length.
8. The method of claim 7 , wherein the brush molecules are from 2-100 nm in length.
9. The method of claim 1 , wherein the first member of the specific binding pair comprises a plurality of different first members of specific binding pairs, the method comprising depositing each different first member of the plurality at a discrete location on the non-fouling polymer layer.
10. The method of claim 7 , wherein the brush molecules have a density of up to 500 milligrams per meter 2 .
11. The method of claim 1 , wherein the first member of the specific binding pair is selected from the group consisting of a protein, a peptide, an antibody, a receptor, a ligand, an antigen, a nucleotide, a synthetic nucleic acid, a carbohydrate, a lipid, a drug, a natural nucleic acid, a cell, and any combination thereof.
12. The method of claim 1 , wherein the first member of the specific binding pair is an antibody.
13. The method of claim 1 , wherein the first member of the specific binding pair is an antigen.
14. The method of claim 11 , wherein the first member of the specific binding pair is a carbohydrate.
15. The method of claim 12 , wherein a cell comprises the antibody.
16. The method of claim 13 , wherein a cell comprises the antigen.
17. The method of claim 14 , wherein a cell comprises the carbohydrate.
18. The method of claim 1 , wherein the biological fluid is selected from the group consisting of blood, blood plasma, serum, saliva, mucus, tear fluid, and urine.
19. The method of claim 18 , wherein the biological fluid is blood.
20. The method of claim 18 , wherein the biological fluid is blood plasma.
21. The method of claim 18 , wherein the biological fluid is saliva.
22. The method of claim 18 , wherein the biological fluid is mucus.
23. The method of claim 18 , wherein the biological fluid is tear fluid.
24. The method of claim 18 , wherein the biological fluid is urine.
25. The method of claim 1 , wherein a second member of the specific binding pair is selected from the group consisting of a protein, a peptide, an antibody, a receptor, a ligand, an antigen, a nucleotide, a synthetic nucleic acid, a carbohydrate, a drug, a blood lipoprotein, and a natural nucleic acid.
26. The method of claim 25 , wherein the second member of the specific binding pair is an antibody.
27. The method of claim 25 , wherein the second member of the specific binding pair is an antigen.
28. The method of claim 25 , wherein the second member of the specific binding pair is a blood lipoprotein.
29. The method of claim 25 , wherein the second member of the specific binding pair is a carbohydrate.
30. The method of claim 26 , wherein a cell comprises the antibody.
31. The method of claim 27 , wherein a cell comprises the antigen.
32. The method of claim 28 , wherein a cell comprises the blood lipoprotein.
33. The method of claim 29 , wherein a cell comprises the carbohydrate.
34. The method of claim 1 , wherein a second member of the specific binding pair is selected from the group consisting of a human cytokine, a human angiogenic factor, a cardiac marker, a diabetes marker, an obesity marker, a fungal infection marker, a viral infection marker, and a microbial infection marker.