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 solid support comprising a plurality of sites, wherein each individual site of the plurality of sites comprises:
i) only one particle coupled to the individual site; and
ii) only one polypeptide of interest attached to the only one particle;
b) delivering binding reagents to the solid support, thereby coupling binding reagents to polypeptides of interest at sites of the plurality of sites;
c) repeating step b) using second binding reagents instead of the binding reagents, wherein the second binding reagents are different from the binding reagents;
d) for each individual site, detecting presence or absence of coupling of each individual binding reagent and second binding reagent to the polypeptide of interest; and
e) based upon presence or absence of coupling of each individual binding reagent and second binding reagent to the polypeptide of interest, characterizing the polypeptide of interest at each individual site of the plurality of sites.
2 . The method of claim 1 , wherein the only one particle is coupled to the individual site by nucleic acid hybridization.
3 . The method of claim 2 , wherein the only one particle comprises one or more oligonucleotides, wherein the individual site comprises one or more complementary oligonucleotides, and wherein the one or more oligonucleotides of the particle are hybridized to the one or more complementary oligonucleotides of the individual site.
4 . The method of claim 1 , wherein the particle comprises a surface-interacting moiety.
5 . The method of claim 4 , wherein the surface-interacting moiety comprises a polymer nanoparticle.
6 . The method of claim 5 , wherein the polymer nanoparticle comprises a nucleic acid nanoparticle.
7 . The method of claim 1 , wherein the binding reagents comprise affinity reagents.
8 . The method of claim 7 , wherein an affinity reagent of the affinity reagents comprises an aptamer.
9 . The method of claim 7 , wherein an affinity reagent of the affinity reagents comprises an antibody or an antibody fragment.
10 . The method of claim 7 , wherein coupling binding reagents to polypeptides of interest comprises coupling an affinity reagent to an epitope of the polypeptide of interest.
11 . The method of claim 7 , wherein coupling binding reagents to polypeptides of interest comprises coupling an affinity reagent to a modified N-terminal amino acid of the polypeptide of interest.
12 . The method of claim 1 , wherein a polypeptide of interest has a full-length primary amino acid structure.
13 . The method of claim 1 , wherein a polypeptide of interest is a fragment of a full-length primary amino acid structure.
14 . The method of claim 1 , wherein providing the solid support comprises providing the solid support comprising a first species of polypeptide and a second species of polypeptide.
15 . The method of claim 14 , wherein the first species of polypeptide and the second species of polypeptide have a dynamic range of at least 10 4 .
16 . The method of claim 15 , wherein characterizing the polypeptide of interest at each individual site of the plurality of sites comprises identifying a polypeptide of the first species of polypeptide and a polypeptide of the second species of polypeptide.
17 . The method of claim 1 , wherein providing the solid support comprises providing the solid support comprising at least 1000 unique polypeptide species of interest.
18 . The method of claim 17 , wherein characterizing the polypeptide of interest at each individual site of the plurality of sites comprises identifying the 1000 unique polypeptide species of interest.
19 . The method of claim 1 , wherein providing the solid support comprises providing the solid support comprising at least 25% of polypeptide species of a proteome.
20 . The method of claim 19 , wherein characterizing the polypeptide of interest at each individual site of the plurality of sites comprises identifying the at least 25% of polypeptide species of the proteome.
21 . The method of claim 1 , wherein detecting presence or absence of coupling of each individual binding reagent to the polypeptide of interest comprises detecting presence or absence of fluorescent signals at an address of the site comprising the polypeptide of interest.
22 . The method of claim 21 , wherein detecting presence or absence of fluorescent signals at an address of the site comprising the polypeptide of interest occurs for each instance of delivering binding reagents to the solid support.
23 . The method of claim 1 , wherein detecting presence or absence of coupling of each individual binding reagent to the polypeptide of interest comprises detecting presence or absence of a nucleic acid tag.
24 . The method of claim 1 , wherein characterizing the polypeptide of interest comprises identifying a species of the polypeptide of interest.
25 . The method of claim 1 , wherein characterizing the polypeptide of interest comprises identifying an amino acid sequence of the polypeptide of interest.
26 . The method of claim 1 , wherein characterizing the polypeptide of interest comprises identifying a proteoform of the polypeptide of interest.
27 . The method of claim 1 , wherein the polypeptide of interest is attached to the particle by a linking moiety.
28 . The method of claim 27 , wherein the linking moiety provides a separation gap of at least 10 nanometers (nm) between the polypeptide of interest and the solid support.
29 . The method of claim 1 , further comprising removing the binding reagents from the polypeptides of interest.
30 . The method of claim 27 , wherein the linking moiety comprises a double-stranded nucleic acid.
31 . The method of claim 27 , wherein the particle comprises a first face and a second face, wherein the first face is attached to the solid support, wherein the second face is oriented away from the solid support, and wherein the second face comprises the linking moiety.
32 . The method of claim 1 , wherein the plurality of sites is provided in a plurality of wells.
33 . The method of claim 32 , wherein each well of the plurality of wells comprises only one site of the plurality of sites.
34 . The method of claim 1 , wherein each binding reagent of the binding reagents binds to a first set of two or more structurally dissimilar polypeptides.
35 . The method of claim 34 , wherein each binding reagent of the second binding reagents binds to a second set of two or more structurally dissimilar polypeptides, wherein the first set of two or more structurally dissimilar polypeptides differs from the second set of two or more structurally dissimilar polypeptides.