IP Library › Granted Patent US 12,305,167
Granted Patent B2
US 12,305,167 · App. 18/770,462 · Granted May 20, 2025

Systems and methods for biomolecule retention

Inventors: Pengyu Hao (Belmont, CA); Elvis Ikwa (San Leandro, CA); Pierre Indermuhle (Berkeley, CA); Christina Inman (San Mateo, CA); Parag Mallick (San Mateo, CA); Torri Elise Rinker (San Francisco, CA); Steven Tan (San Mateo, CA); Tural Aksel (Redwood City, CA); Stephen Hendricks (Los Gatos, CA); Sadie Ingle (Suttons Bay, MI)
Assignee: Nautilus Subsidiary, Inc.
C12N15/1093B01J19/0046B82Y5/00C12Q1/6804C12Q1/6837B01J2219/0061
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Quick Facts
Patent No.
US 12,305,167
App. No.
18/770,462
Granted
May 20, 2025
Kind
B2
Abstract

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.

Claims (47)

1. A method, comprising:

a) forming a first plurality of nucleic acid nanoparticles, wherein the first plurality of nucleic acid nanoparticles is attached to a first plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles is attached to only one polypeptide of the first plurality of polypeptides of interest, wherein a first unique identifier is attached to each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles, and wherein the first plurality of polypeptides of interest comprises at least 10 3 different native-length protein primary sequences;

b) forming a second plurality of nucleic acid nanoparticles, wherein the second plurality of nucleic acid nanoparticles is attached to a second plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles is attached to only one polypeptide of the second plurality of polypeptides of interest, wherein a second unique identifier is attached to each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles, wherein the first unique identifier and the second unique identifier are different, and wherein the second plurality of polypeptides of interest comprises at least 10 3 different native-length protein primary sequences;

c) attaching to each site of a plurality of sites of a solid support only one nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles or the second plurality of nucleic acid nanoparticles, wherein each site of the plurality of sites is optically resolvable from any other site of the plurality of sites; and

d) detecting signals from the first unique identifiers at a first set of sites of the plurality of sites, detecting signals from the second unique identifiers at a second set of sites of the plurality of sites.

2. The method of claim 1 , further comprising obtaining the first plurality of polypeptides of interest from a first sample.

3. The method of claim 2 , further comprising obtaining the second plurality of polypeptides of interest from a second sample.

4. The method of claim 2 , further comprising obtaining the second plurality of polypeptides of interest from the first sample.

5. The method of claim 1 , wherein attaching to the plurality of sites of the solid support the first plurality of nucleic acid nanoparticles and the second plurality of nucleic acid nanoparticles comprises attaching at a site of the first set of sites a pendant oligonucleotide of a nucleic acid nanoparticle of the first plurality of nanoparticles to a surface-coupled oligonucleotide of the site.

6. The method of claim 1 , wherein attaching to the plurality of sites of the solid support the first plurality of nucleic acid nanoparticles and the second plurality of nucleic acid nanoparticles comprises attaching at a site of the second set of sites a pendant oligonucleotide of a nucleic acid nanoparticle of the second plurality of nanoparticles to a surface-coupled oligonucleotide of the site.

7. The method of claim 1 , wherein each site of the plurality of sites comprises one or more surface-coupled oligonucleotides, wherein each oligonucleotide of the one or more surface-coupled oligonucleotides comprises an identical nucleotide sequence.

8. The method of claim 7 , wherein each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles comprises a pendant oligonucleotide, wherein the pendant oligonucleotide comprises a complementary nucleotide sequence to the identical nucleotide sequence.

9. The method of claim 8 , wherein each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles comprises a pendant oligonucleotide, wherein the pendant oligonucleotide comprises the complementary nucleotide sequence to the identical nucleotide sequence.

10. The method of claim 9 , further comprising hybridizing the complementary nucleotide sequence of the pendant oligonucleotide of a nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles to the identical nucleotide sequence of a site of the first set of sites of the plurality of sites.

11. The method of claim 10 , further comprising hybridizing the complementary nucleotide sequence of the pendant oligonucleotide of a nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles to the identical nucleotide sequence of a site of the second set of sites of the plurality of sites.

12. The method of claim 1 , wherein attaching to the plurality of sites of the solid support the first plurality of nucleic acid nanoparticles and the second plurality of nucleic acid nanoparticles comprises:

i) attaching the first plurality of nucleic acid nanoparticles to the first set of sites; and

ii) after attaching the first plurality of nucleic acid nanoparticles to the first set of sites, attaching the second plurality of nucleic acid nanoparticles to the second set of sites.

13. The method of claim 12 , further comprising, before attaching the second plurality of nucleic acid nanoparticles to the second set of sites, determining the first set of sites containing the first plurality of nucleic acid nanoparticles.

14. The method of claim 13 , wherein determining the first set of sites containing the first plurality of nucleic acid nanoparticles comprises detecting signals from nucleic acid nanoparticles at addresses of the first set of sites.

15. The method of claim 12 , further comprising, after attaching the second plurality of nucleic acid nanoparticles to the second set of sites, determining the second set of sites containing the second plurality of nucleic acid nanoparticles.

16. The method of claim 15 , wherein determining the second set of sites containing the second plurality of nucleic acid nanoparticles comprises detecting signals from nucleic acid nanoparticles at addresses of the second set of sites.

17. The method of claim 1 , wherein the first unique identifier comprises a first detectable label and the second unique identifier comprises a second detectable label, wherein the first detectable label differs from the second detectable label.

18. The method of claim 17 , wherein the first detectable label and the second detectable label comprise differing fluorophores.

19. The method of claim 17 , wherein the first detectable label and the second detectable label comprises differing nucleic acid tags or peptide tags.

20. The method of claim 1 , wherein the first set of sites and the second set of sites have random spatial distributions.

21. The method of claim 1 , wherein the attaching occurs in the presence of a fluidic medium containing a denaturing species.

22. The method of claim 1 , wherein the first plurality of polypeptides of interest or the second plurality of polypeptides of interest comprises at least 10 6 polypeptide molecules.

23. The method of claim 22 , wherein the first plurality of polypeptides of interest comprises at least 10 6 polypeptide molecules and the second plurality of polypeptides of interest comprises at least 10 6 polypeptide molecules.

24. The method of claim 22 , wherein the first plurality of polypeptides of interest or the second plurality of polypeptides of interest comprises at least 10 8 polypeptide molecules.

25. The method of claim 1 , wherein the plurality of sites comprises at least 10 8 sites.

26. The method of claim 1 , wherein a site of the plurality of sites is separated by at least 500 nm from any other site of the plurality of sites.

27. A method, comprising:

a) forming a first plurality of nucleic acid nanoparticles, wherein the first plurality of nucleic acid nanoparticles is attached to a first plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles is attached to only one polypeptide of the first plurality of polypeptides of interest, wherein a first unique identifier is attached to each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles, and wherein the first plurality of polypeptides comprises at least 25% of polypeptide species of a proteome;

b) forming a second plurality of nucleic acid nanoparticles, wherein the second plurality of nucleic acid nanoparticles is attached to a second plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles is attached to only one polypeptide of the second plurality of polypeptides of interest, wherein a second unique identifier is attached to each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles, wherein the first unique identifier and the second unique identifier are different, and wherein the second plurality of polypeptides comprises at least 25% of polypeptide species of a proteome;

c) attaching to each site of a plurality of sites of a solid support only one nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles or the second plurality of nucleic acid nanoparticles, wherein each site of the plurality of sites is optically resolvable from any other site of the plurality of sites; and

d) detecting signals from the first unique identifiers at a first set of sites of the plurality of sites, and detecting signals from the second unique identifiers at a second set of sites of the plurality of sites.

28. A method, comprising:

a) forming a first plurality of nucleic acid nanoparticles, wherein the first plurality of nucleic acid nanoparticles is attached to a first plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles is attached to only one polypeptide of the first plurality of polypeptides of interest, wherein a first unique identifier is attached to each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles, and wherein the first plurality of polypeptides comprises a complexity of at least 10 5 different protein structures;

b) forming a second plurality of nucleic acid nanoparticles, wherein the second plurality of nucleic acid nanoparticles is attached to a second plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles is attached to only one polypeptide of the second plurality of polypeptides of interest, wherein a second unique identifier is attached to each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles, wherein the first unique identifier and the second unique identifier are different, and wherein the second plurality of polypeptides comprises a complexity of at least 10 5 different protein structures;

c) attaching to each site of a plurality of sites of a solid support only one nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles or the second plurality of nucleic acid nanoparticles, wherein each site of the plurality of sites is optically resolvable from any other site of the plurality of sites; and

d) detecting signals from the first unique identifiers at a first set of sites of the plurality of sites, and detecting signals from the second unique identifiers at a second set of sites of the plurality of sites.

29. A method, comprising:

a) forming a first plurality of nucleic acid nanoparticles, wherein the first plurality of nucleic acid nanoparticles is attached to a first plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles is attached to only one polypeptide of the first plurality of polypeptides of interest, wherein a first unique identifier is attached to each nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles, and wherein the first plurality of polypeptides comprises a dynamic range of at least 10 4 ;

b) forming a second plurality of nucleic acid nanoparticles, wherein the second plurality of nucleic acid nanoparticles is attached to a second plurality of polypeptides of interest, wherein each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles is attached to only one polypeptide of the second plurality of polypeptides of interest, wherein a second unique identifier is attached to each nucleic acid nanoparticle of the second plurality of nucleic acid nanoparticles, wherein the first unique identifier and the second unique identifier are different, and wherein the second plurality of polypeptides comprises a dynamic range of at least 10 4 ;

c) attaching to each site of a plurality of sites of a solid support only one nucleic acid nanoparticle of the first plurality of nucleic acid nanoparticles or the second plurality of nucleic acid nanoparticles, wherein each site of the plurality of sites is optically resolvable from any other site of the plurality of sites; and

d) detecting signals from the first unique identifiers at a first set of sites of the plurality of sites, and detecting signals from the second unique identifiers at a second set of sites of the plurality of sites.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 5, 2024
From: HAO, PENGYU; IKWA, ELVIS; INDERMUHLE, PIERRE; INMAN, CHRISTINA; MALLICK, PARAG; RINKER, TORRI ELISE; TAN, STEVEN; AKSEL, TURAL; HENDRICKS, STEPHEN; INGLE, SADIE
To: NAUTILUS SUBSIDIARY, INC.
Reel/Frame 069501/0722 →
Continuity (7)
Continuation 18416639 · Jan 18, 2024
Continuation 18361731 · Jul 28, 2023
Continuation 18050732 · Oct 28, 2022
Continuation 17692035 · Mar 10, 2022
Provisional Application 63256761 · Oct 18, 2021
Provisional Application 63159500 · Mar 11, 2021
Related Publication 20240368585A1 · Nov 7, 2024
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