Systems and methods for biomolecule retention
Compositions, systems, and methods for the display of analytes such as biomolecules are described. Display of analytes is achieved by coupling of the analytes to displaying molecules that are configured to associate with surfaces or interfaces. Arrays of analytes may be formed from the described systems for utilization in assays and other methods.
1. A method, comprising:
a) providing a plurality of nucleic acid nanostructures, wherein each nucleic acid nanostructure is coupled to one and only one polypeptide by a separating group, wherein the separating group provides an average separation distance of at least 10 nanometers between the one and only one polypeptide and a nucleic acid nanostructure to which the one and only polypeptide is coupled;
b) coupling the plurality of nucleic acid nanostructures to a plurality of sites of an array, wherein the plurality of nucleic acid nanostructures comprises a first polypeptide species and a second polypeptide species having a dynamic range of at least 10 4 ; and
c) detecting presence of the first polypeptide species and the second polypeptide species at individual sites of the array.
2. The method of claim 1 , further comprising: i) coupling affinity agents to polypeptides at the sites of the plurality of sites, and ii) detecting presence or absence of an affinity agent at each site of the plurality of sites.
3. The method of claim 2 , wherein detecting the presence or absence of the affinity agent at each site of the plurality of sites comprises detecting presence or absence of a signal from a detectable label coupled to the affinity agent.
4. The method of claim 2 , further comprising identifying a polypeptide of the first polypeptide species and a polypeptide of the second polypeptide species based upon detected presence or absence of affinity agents at each site of the plurality of sites.
5. The method of claim 1 , wherein forming the plurality of nucleic acid nanostructures comprises coupling a plurality of polypeptides to a plurality of nucleic acid nanostructure components.
6. The method of claim 5 , wherein nucleic acid nanostructures of the plurality of nucleic acid nanostructures comprise linking moieties, and wherein coupling the plurality of polypeptides to the plurality of nucleic acid nanostructures comprises coupling polypeptides to the linking moieties.
7. The method of claim 1 , further comprising detecting a terminal amino acid residue of a polypeptide of the first polypeptide species or the second polypeptide species on the array.
8. The method of claim 1 , wherein coupling a nucleic acid nanostructure of the plurality of nucleic acid nanostructures to a site of the plurality of sites comprises hybridizing a nucleic acid strand of the nucleic acid nanostructure to a complementary nucleic acid strand, wherein the complementary nucleic acid strand is coupled to the site.
9. The method of claim 1 , wherein a nucleic acid nanostructure of the plurality of nucleic acid nanostructures comprises a nucleic acid origami.
10. The method of claim 9 , wherein the nucleic acid nanostructure of the plurality of nucleic acid nanostructures further comprises a pervious structure, wherein the pervious structure comprises a plurality of pendant moieties.
11. The method of claim 10 , wherein coupling the nucleic acid nanostructure of the plurality of nucleic acid nanostructures to the site of the plurality of sites comprises coupling a pendant moiety of the plurality of pendant moieties to the site of the plurality of sites.
12. The method of claim 11 , wherein coupling the nucleic acid nanostructure of the plurality of nucleic acid nanostructures to the site of the plurality of sites comprises coupling a subset of pendant moieties of the plurality of pendant moieties to the site of the plurality of sites.
13. The method of claim 11 , wherein the pendant moiety of the plurality of pendant moieties comprises a single-stranded nucleic acid.
14. The method of claim 11 , wherein the pendant moiety of the plurality of pendant moieties comprises a polymer chain.
15. The method of claim 1 , further comprising: i) fragmenting a polypeptide of the first polypeptide species or the second polypeptide species into at least one peptide fragment, and ii) coupling a peptide fragment of the at least one peptide fragment to a nucleic acid nanostructure.
16. The method of claim 1 , wherein polypeptides of the array comprise a complexity of at least about 1×10 3 native-length primary protein sequences.
17. The method of claim 1 , wherein the separating group comprises a polymer linker, a nucleic acid linker, or a nanoparticle linker.
18. The method of claim 17 , wherein the polymer linker comprises a polyethylene glycol (PEG) moiety.
19. The method of claim 17 , wherein the nucleic acid linker comprises a double-stranded nucleic acid.
20. The method of claim 1 , wherein the separating group provides an average separation distance of at least 15 nanometers between the one and only one polypeptide and a surface of a solid support.
21. The method of claim 1 , wherein detecting presence of the first polypeptide species and the second polypeptide species at individual sites of the array comprises detecting presence of a plurality of polypeptides of the first polypeptide species.
22. The method of claim 21 , wherein detecting presence of the first polypeptide species and the second polypeptide species at individual sites of the array comprises detecting presence of a plurality of polypeptides of the second polypeptide species.
23. The method of claim 22 , further comprising determining a quantity of polypeptides of the first polypeptide species and a quantity of polypeptides of the second polypeptide species.
24. The method of claim 23 , further comprising characterizing the dynamic range of the first polypeptide species and the second polypeptide species based upon the quantity of polypeptides of the first polypeptide species and the quantity of polypeptides of the second polypeptide species.
25. The method of claim 1 , wherein providing the plurality of nucleic acid nanostructures comprises coupling polypeptides to separating groups, thereby forming the plurality of nucleic acid nanostructures.
26. The method of claim 25 , wherein coupling polypeptides to separating groups comprises covalently coupling polypeptides to separating groups.
27. The method of claim 25 , further comprising purifying the plurality of nucleic acid nanostructures.
28. The method of claim 27 , wherein purifying the plurality of nucleic acid nanostructures comprises separating from the plurality of nucleic acid nanostructures a nucleic acid nanostructure that is not coupled to a polypeptide.
29. The method of claim 27 , wherein purifying the plurality of nucleic acid nanostructures comprises separating from the plurality of nucleic acid nanostructures a nucleic acid nanostructure that is coupled to two or more polypeptides.