# US 2024/0201182 A1 · Inhibition of photon phenomena on single molecule arrays

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- Number: US 2024/0201182 A1
- Document ID: US20240201182A1
- Status: Published application
- Kind: A1
- Application number: 18/540,682
- Filed: 2023-12-14
- Published: 2024-06-20
- Earliest claimed: 2022-12-15
- Assignees: Nautilus Subsidiary, Inc.
- Applicants: Nautilus Subsidiary, Inc.
- Inventors: Seok Ki CHOI; Michael Augusto DARCY; Steven TAN; Pengyu HAO
- Web page: https://patentgrep.com/patent/US20240201182A1

## CPC

- G01N 33/54346: {Nanoparticles}
- G01N 33/54386: {Analytical elements}
- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G01N 33/54373: {involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings}
- G01N 33/54393: {Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding}
- G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates}
- G01N 21/6456: {Spatial resolved fluorescence measurements; Imaging}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Methods of providing photodamage inhibitors to single-analyte arrays are described. Photodamage inhibitors may be incorporated into various reagents that are subsequently coupled to single-analyte arrays. Photodamage inhibitors may be replenished during a single-analyte assay through provision of more reagents containing photodamage inhibitors. Useful reagent compositions containing photodamage inhibitors are described.

## Claims

### Claim 1

A method, comprising:
a) providing a single-analyte array comprising a plurality of sites, wherein individual sites of the plurality of sites each comprise one and only one macromolecule of a first plurality of macromolecules;
b) binding a second plurality of macromolecules to macromolecules of the first plurality of macromolecules at a fraction of sites of the plurality of sites, thereby forming a macromolecule complex at respective sites of the fraction of sites, wherein the macromolecule complex comprises a macromolecule of the second plurality of macromolecules bound to one and only one macromolecule of the first plurality of macromolecules;
c) detecting, in the presence of photons, a signal at each individual site of the first fraction of sites; and
d) after detecting, in the presence of photons, the signal at respective sites of the first fraction of sites, dissociating macromolecules of the second plurality of macromolecules from macromolecules of the first plurality of macromolecules;
wherein individual macromolecule complexes each comprise a plurality of photodamage inhibitor moieties that are coupled to the individual macromolecule complex.

### Claim 2 (depends on 1)

The method of claim 1, wherein a macromolecule of the first plurality of macromolecules comprises an analyte of interest.

### Claim 3 (depends on 2)

The method of claim 2, wherein the analyte of interest is coupled to an anchoring moiety.

### Claim 4 (depends on 3)

The method of claim 3, wherein the anchoring moiety comprises a nanoparticle.

### Claim 5 (depends on 1)

The method of claim 1, wherein a macromolecule of the second plurality of macromolecules comprises an affinity agent.

### Claim 6 (depends on 5)

The method of claim 5, wherein the macromolecule of the second plurality of macromolecules comprises two or more affinity agents.

### Claim 7 (depends on 5)

The method of claim 5, wherein the macromolecule of the second plurality of macromolecules further comprises a nanoparticle.

### Claim 8 (depends on 7)

The method of claim 7, wherein the nanoparticle comprises a polymer nanoparticle.

### Claim 9 (depends on 8)

The method of claim 8, wherein the polymer nanoparticle comprises a nucleic acid nanoparticle.

### Claim 10 (depends on 8)

The method of claim 8, wherein the polymer nanoparticle comprises a branched or dendrimeric polymer nanoparticle.

### Claim 11 (depends on 1)

The method of claim 1, wherein a photodamage inhibitor moiety of the plurality of photodamage inhibitor moieties is coupled to a macromolecule of the first plurality of macromolecules.

### Claim 12 (depends on 1)

The method of claim 1, wherein a photodamage inhibitor moiety of the plurality of photodamage inhibitor moieties is coupled to a macromolecule of the second plurality of macromolecules.

### Claim 13 (depends on 1)

The method of claim 1, wherein a photodamage inhibitor moiety of the plurality of photodamage inhibitor moieties is coupled to a surface-coupled moiety of a site of the fraction of sites.

### Claim 14 (depends on 1)

The method of claim 1, wherein binding the second plurality of macromolecules to macromolecules of the first plurality of macromolecules at the fraction of sites of the plurality of sites comprises binding an avidity component of a macromolecule of the second plurality of macromolecules to a complementary avidity component of a site of the fraction of sites.

### Claim 15 (depends on 14)

The method of claim 14, wherein a photodamage inhibitor moiety of the plurality of photodamage inhibitor moieties is coupled to the avidity component of the macromolecule of the second plurality of macromolecules.

### Claim 16 (depends on 15)

The method of claim 15, wherein a photodamage inhibitor moiety of the plurality of photodamage inhibitor moieties is coupled to the complementary avidity component of the site of the fraction of sites.

### Claim 17 (depends on 1)

The method of claim 1, wherein dissociating macromolecules of the second plurality of macromolecules from macromolecules of the first plurality of macromolecules comprises dissociating macromolecules of the second plurality of macromolecules from macromolecules of the first plurality of macromolecules at at least 95% of sites of the fraction of sites.

### Claim 18 (depends on 1)

The method of claim 1, wherein the detecting, in the presence of photons, the signal at each individual site of the first fraction of sites occurs in the presence of a fluidic medium.

### Claim 19 (depends on 18)

The method of claim 18, wherein the fluidic medium comprises a second plurality of photodamage inhibitors.

### Claim 20

An array composition, comprising:
a) a plurality of sites, wherein individual sites of the plurality of sites each comprise one and only one macromolecule of a first plurality of macromolecules;
b) at individual sites of a fraction of sites of the plurality of sites, a macromolecule of a second plurality of macromolecules bound to the one and only one macromolecule of the first plurality of macromolecules; and
c) at individual sites of the fraction of sites of the plurality of sites, a plurality of photodamage inhibitor moieties coupled to a respective individual site;
wherein the fraction of sites contains an average of at least 100 photodamage inhibitor moieties per site.