Compositions, systems, and methods for detecting the presence of polymer subunits using chemiluminescence

Patent metadata

Field	Value
Publication number	US-11629373-B2
Application number	US-202016832309-A
Country	US
Kind code	B2
Filing date	Mar 27, 2020
Priority date	Sep 12, 2014
Publication date	Apr 18, 2023
Grant date	Apr 18, 2023

Abstract

Under one aspect, a composition includes a substrate; a first polynucleotide coupled to the substrate; a second polynucleotide hybridized to the first polynucleotide; and a catalyst coupled to a first nucleotide of the second polynucleotide, the catalyst being operable to cause a chemiluminogenic molecule to emit a photon. Under another aspect, a method includes providing a catalyst operable to cause a first chemiluminogenic molecule to emit a photon; providing a substrate; providing a first polynucleotide coupled to the substrate; hybridizing a second polynucleotide to the first polynucleotide; coupling a first quencher to a first nucleotide of the second polynucleotide; and inhibiting, by the first quencher, photon emission by the first chemiluminogenic molecule.

First claim

What is claimed is:

1. A composition comprising:
a catalyst operable to cause a chemiluminogenic molecule to emit a photon;
a substrate;
a first polynucleotide coupled to the substrate;
a second polynucleotide hybridized to the first polynucleotide; and
a quencher coupled to a first nucleotide at a 3′ end of the second polynucleotide, the quencher operable to inhibit photon emission by the chemiluminogenic molecule.

2. The composition of claim 1 , wherein the catalyst is coupled to the substrate or coupled to the first polynucleotide.

3. The composition of claim 1 , wherein the quencher is cleavable from the first nucleotide.

4. The composition of claim 1 , wherein the quencher is coupled to the first nucleotide via a first moiety, and a second moiety is coupled to the first moiety and to the quencher.

5. The composition of claim 1 , wherein the catalyst is selected from the group consisting of an enzyme, a metallic catalyst, and a metalorganic catalyst and wherein the enzyme is selected from the group consisting of: a luciferase, a 1,2-dioxetane cleaver and a peroxide generator.

6. The composition of claim 1 wherein the chemiluminogenic molecule is selected from the group consisting of a luciferin, a luciferin derivative, coelenterazine, a coelenterazine derivative, a 1,2-dioxetane derivative, luminol, a luminol derivative, acridinium, and an acridinium derivative.

7. The composition of claim 1 , wherein the quencher is selected from the group consisting of a DABCYL quencher, a BHQ quencher, (±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, 2,4-dinitrophenol (2,4-DNP), 2,5-dinitrophenol (2,5-DNP), and 2,6-dinitrophenol (2,6-DNP).

8. A system comprising: the composition of claim 1 , and circuitry configured to detect a photon emitted by the chemiluminogenic molecule.

9. A method comprising:
obtaining the composition of claim 1 by:
providing a catalyst operable to cause a first chemiluminogenic molecule to emit a photon;
providing a substrate;
providing a first polynucleotide coupled to the substrate;
hybridizing a second polynucleotide to the first polynucleotide; and
coupling a first quencher to a first nucleotide at the 3′ end of the second polynucleotide, wherein the first quencher inhibits photon emission by the first chemiluminogenic molecule.

10. The method of claim 9 , further comprising:
cleaving the first quencher from the first nucleotide;
adding a second nucleotide to the 3′ end of second polynucleotide; and
coupling a second quencher to the second nucleotide.

11. The method of claim 10 , wherein the second nucleotide is coupled to a first moiety, the second quencher is coupled to a second moiety, wherein said second quencher is coupled to the second nucleotide by coupling the second moiety to the first moiety, and wherein the second quencher inhibits photon emission by a second chemiluminogenic molecule;
detecting a photon emitted by the second chemiluminogenic molecule in the absence of the second quencher;
detecting the presence of the second nucleotide based on inhibition of emission of the photon by the second chemiluminogenic molecule;
detecting a photon emitted by the first chemiluminogenic molecule in the absence of the first quencher; and
detecting the presence of the first nucleotide based on detection of inhibition of emission of the photon by the first chemiluminogenic molecule.

12. The composition of claim 1 , further comprising a plurality of the chemiluminogenic molecules, wherein the quencher inhibits photon emission by each of the chemiluminogenic molecules.

13. The composition of claim 12 , further comprising a plurality of reagent molecules, wherein the catalyst is capable of causing each of the chemiluminogenic molecules to emit a corresponding photon by oxidizing the chemiluminogenic molecule using a reagent molecule in the absence of the quencher to form an oxidized chemiluminogenic molecule.

14. The composition of claim 13 , wherein the oxidized chemiluminogenic molecule has an excited state that decays by emitting the corresponding photon in the absence of the quencher.

15. The composition of claim 4 , wherein one of the first and second moieties is biotin or a biotin derivative, and wherein the other of the first and second moieties is streptavidin.

16. The composition of claim 15 , wherein one of the first and second moieties is digoxigenin, and wherein the other of the first and second moieties is anti-digoxigenin.

17. The composition of claim 1 , wherein the catalyst comprises a peroxide generator, and wherein the chemiluminogenic molecule comprises luminol, a luminol derivative, acridinium, or an acridinium derivative.

18. The composition of claim 17 , wherein the chemiluminogenic molecule comprises luminol, or a luminol derivative.

19. The composition of claim 17 , wherein the chemiluminogenic molecule comprises acridinium, or an acridinium derivative.

20. The composition of claim 17 , wherein the peroxide generator comprises an enzyme.

21. The composition of claim 17 , wherein the peroxide generator comprises a metallic catalyst, an organic catalyst, or a metalorganic catalyst.

Assignees

• Illumina Inc

Inventors

• Boyanov Boyan
• Qiang Liangliang
• Gunderson Kevin L
• Klausing Kay
• Pickering Lea
• Delattre Cyril
• Khurana Tarun

Classifications

• C12Q1/6818 Primary

  involving interaction of two or more labels, e.g. resonant energy transfer · CPC title

• C12Q2563/125

  the label being enzymatic, i.e. proteins, and non proteins, such as nucleic acid with enzymatic activity · CPC title

• C12Q2535/122

  Massive parallel sequencing · CPC title

• C12Q2563/103

  luminescence · CPC title

• C12Q2533/101

  Primer extension · CPC title