Functionalized nanostructures and flow cell depressions

Patent metadata

Field	Value
Publication number	US-2023348970-A1
Application number	US-202318176761-A
Country	US
Kind code	A1
Filing date	Mar 1, 2023
Priority date	Mar 1, 2022
Publication date	Nov 2, 2023
Grant date	—

Abstract

A functionalized nanostructure includes a metal nanostructure; an un-cleavable first primer and a cleavable second primer attached to a first region of the metal nanostructure through i) a first thiol linkage attached to a first polymer chain having a first polarity or ii) respective first thiol linkages attached to respective first polymer chains having the first polarity; and a cleavable first primer and an un-cleavable second primer attached to a second region of the metal nanostructure through i) a second thiol linkage attached to a second polymer chain having a second polarity different from the first polarity or ii) respective second thiol linkages attached to respective second polymer chains having the second polarity.

First claim

1 . A functionalized nanostructure, comprising:
a metal nanostructure; an un-cleavable first primer and a cleavable second primer attached to a first region of the metal nanostructure through i) a first thiol linkage attached to a first polymer chain having a first polarity or ii) respective first thiol linkages attached to respective first polymer chains having the first polarity; and a cleavable first primer and an un-cleavable second primer attached to a second region of the metal nanostructure through i) a second thiol linkage attached to a second polymer chain having a second polarity different from the first polarity or ii) respective second thiol linkages attached to respective second polymer chains having the second polarity.

2 . The functionalized nanostructure as defined in claim 1 , wherein the metal nanostructure is selected from the group consisting of gold, platinum, copper, and silver.

3 . The functionalized nanostructure as defined in claim 1 , wherein:
the un-cleavable first primer and the cleavable second primer are attached to the first region through the respective first thiol linkages attached to the respective first polymer chains; the un-cleavable first primer and the cleavable second primer are attached to the respective first polymer chains at respective ends that are opposed to the respective first thiol linkages; the cleavable first primer and the un-cleavable second primer are attached to the second region through the respective second thiol linkages attached to the respective second polymer chains; and the cleavable first primer and the un-cleavable second primer are attached to the respective second polymer chains at respective ends that are opposed to the respective second thiol linkages.

4 . The functionalized nanostructure as defined in claim 3 , wherein:
each of the respective first polymer chains is polystyrene; and each of the respective second polymer chains is poly(ethylene glycol).

5 . The functionalized nanostructure as defined in claim 1 , wherein:
the un-cleavable first primer and the cleavable second primer are attached to the first region through the first thiol linkage attached to the first polymer chain; the un-cleavable first primer and the cleavable second primer are attached to respective side chains of the first polymer chain; the cleavable first primer and the un-cleavable second primer are attached to the second region through the second thiol linkage attached to the second polymer chain; and the cleavable first primer and the un-cleavable second primer are attached to the respective side chains of the second polymer chain.

6 . The functionalized nanostructure as defined in claim 5 , wherein:
each of the respective side chains of the first polymer chain is positioned closer to an end of the first polymer chain that is opposed to the first thiol linkage than to the first thiol linkage; and each of the respective side chains of the second polymer chain is positioned closer to an end of the second polymer chain that is opposed to the second thiol linkage than to the second thiol linkage.

7 . The functionalized nanostructure as defined in claim 6 , wherein:
a predetermined portion of a backbone of the first polymer chain that is adjacent to the first thiol linkage is free of the respective side chains of the first polymer chain; and a predetermined portion of a backbone of second first polymer chain that is adjacent to the second thiol linkage is free of the respective side chains of the first polymer chain.

8 . The functionalized nanostructure as defined in claim 5 , wherein:
the respective side chains of the first polymer chain are distributed along a backbone of the first polymer chain; and the respective side chains of the second polymer chain are distributed along a backbone of the second polymer chain.

9 . A method, comprising:
generating a first functionalized primer set by attaching an un-cleavable first primer and a cleavable second primer to i) a first polymer chain having a first polarity and a first thiol linkage or ii) respective first polymer chains having the first polarity and respective first thiol linkages; generating a second functionalized primer set by attaching each of a cleavable first primer and an un-cleavable second primer to i) a second polymer chain having a second polarity different from the first polarity and a second thiol linkage or ii) respective second polymer chains having the second polarity and respective second thiol linkages; and attaching the first and second functionalized primer sets, through the first thiol linkage and the second thiol linkage or the respective first thiol linkages and the respective second thiol linkages, to spatially separate regions of a metal surface.

10 . The method as defined in claim 9 , wherein:
generating the first functionalized primer set involves reacting respective terminal end functional groups of the un-cleavable first primer and of the cleavable second primer with respective ends groups of the respective first polymer chains; and generating the second functionalized primer set involves reacting respective terminal end functional groups of the cleavable first primer and of the un-cleavable second primer with respective ends groups of the respective second polymer chains.

11 . The method as defined in claim 10 , wherein:
the first functionalized primer set and the second functionalized primer set are generated in separate reaction containers; and the method further comprises:
mixing the first functionalized primer set and the second functionalized primer set to form a mixture; and
exposing the mixture to the metal surface.

12 . The method as defined in claim 11 , wherein one of:
the metal surface includes metal nanostructures, and exposing the mixture to the metal surface involves adding the metal nanostructures to the mixture; or the metal surface includes a transparent metal film positioned on a bottom surface in each of a plurality of depressions of a flow cell, and exposing the mixture to the metal surface involves incubating the mixture in each of the plurality of depressions.

13 . The method as defined in claim 9 , wherein:
generating the first functionalized primer set involves:
polymerizing a first primer grafting monomer and a first backbone monomer to introduce primer reactive side chains to the first polymer chain; and
respectively grafting the un-cleavable first primer and the cleavable second primer to the primer reactive side chains of the first polymer chain; and
generating the second functionalized primer set involves:
polymerizing a second primer grafting monomer and a second backbone monomer to introduce primer reactive side chains to the second polymer chain; and
respectively grafting the cleavable first primer and the un-cleavable second primer to the primer reactive side chains of the second polymer chain.

14 . The method as defined in claim 13 , wherein:
the first functionalized primer set and the second functionalized primer set are generated in separate reaction containers; and the method further comprises:
mixing the first functionalized primer set and the second functionalized primer set to form a mixture; and
exposing the mixture to the metal surface.

15 . The method as defined in claim 14 , wherein one of:
the metal surface includes metal nanostructures, and exposing the mixture to the metal surface involves adding the metal nanostructures to the mixture; or the metal surface includes a transparent metal film positioned on a bottom surface in each of a plurality of depressions of a flow c

Assignees

• Illumina Cambridge Ltd

Inventors

• Artioli Gianluca Andrea
• Redivo Luca
• Richez Alexandre
• Von Hatten Xavier

Classifications

• C12Q1/6874 Primary

  involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• C08F292/00

  Macromolecular compounds obtained by polymerising monomers on to inorganic materials · CPC title

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title