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Reversible surface functionalization

A technology for functionalized, substrate surfaces, applied in the field of chemistry, can solve the problem of not having effective renewal flow cells

Active Publication Date: 2017-08-18
ILLUMINA CAMBRIDGE LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

No effective method of updating the flow cell is commercially available

Method used

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  • Reversible surface functionalization
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  • Reversible surface functionalization

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Experimental program
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Embodiment 1

[0207] figure 2 Embodiments of methods for reversibly attaching biomolecules to silane-functionalized surfaces are shown. First, a substrate comprising a glass surface is treated by chemical vapor deposition (CVD) with a functionalized silane comprising a hydrophobic moiety. Cyclodextrins are used as host molecules, and functionalized primers are grafted onto the cyclodextrins by reacting with one or more azido groups of the cyclodextrins. The silanized surface is then contacted with a cyclodextrin-bonded primer such that the hydrophobic head of the silane forms a host-guest inclusion complex with the lumen of the cyclodextrin via reversible interactions. As a result, the primers are immobilized on the surface of the substrate. Following standard clustering and sequencing procedures, the cyclodextrins are dissociated from the surface by the addition of 1,8-octanediol, a molecule with greater affinity for the cyclodextrin lumen than the hydrophobic head of the functionalized...

Embodiment 2

[0209] In the first experiment, fluorescein isothiocyanate (FITC)-functionalized cyclodextrins were reacted with norbornenesilane-functionalized surfaces at 50 °C for 30 min, followed by rinsing with different solvents. Methods for preparing such norbornene silane functionalized surfaces are described in detail in US Patent Application Serial No. 14 / 316,478, which is incorporated herein by reference in its entirety.

[0210] In this experiment, the standard norbornene silane [(5-bicyclo[2.2.1]hept-2-enyl)ethyl]trimethoxysilane was used to silanize with flow pool. The norbornene moiety is lipophilic and can function as a guest for cyclodextrins. According to molecular mechanical predictions obtained from the literature, only the bridging methylene group from norbornene will fit into the inner cavity of the cyclodextrin (binding free energy of about -2 kcal / mol). Other possible guests with more stable host-guest complexes can also be used, for example guests comprising plan...

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Abstract

Some embodiments described herein relate to a substrate comprising a silane functionalized surface for reversibly immobilizing a biological molecule of interest, such as oligonucleotides, polynucleotides, or protein. Methods for immobilizing the biological molecule and the use in DNA sequencing and other diagnostic applications are also disclosed.

Description

[0001] Incorporation by reference of any priority application [0002] This application claims the benefit of priority from US Provisional Patent Application No. 62 / 040,323, filed August 21, 2014, the entire contents of which are expressly incorporated herein by reference. [0003] field of invention [0004] In general, this application relates to the fields of chemistry, biology, and materials science. More specifically, the present application relates to substrates comprising silane-functionalized surfaces for reversible immobilization of biomolecules of interest. Also disclosed are methods for immobilizing biomolecules and use in DNA sequencing and other diagnostic applications. [0005] Background of the invention [0006] Substrates coated with polymers or hydrogels are used in many technical applications. For example, polymer or hydrogel coated substrates are used for the preparation and / or analysis of biomolecules. Molecular analysis, such as certain nucleic acid se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/551
CPCG01N33/543G01N33/54353G01N33/551C12Q1/6869C12Q1/6834C12Q2563/131C12Q2565/543C12N11/10C09D183/04G01N33/54393
Inventor X.冯海坦W.N.乔治A.里奇兹A-C.丁沃尔A.A.布朗
Owner ILLUMINA CAMBRIDGE LTD