Sample preparation and flow-through sensors using functionalized silicon nanomembranes

Patent metadata

Field	Value
Publication number	US-2020333311-A1
Application number	US-201916959869-A
Country	US
Kind code	A1
Filing date	Jan 7, 2019
Priority date	Jan 5, 2018
Publication date	Oct 22, 2020
Grant date	—

Abstract

Provided are methods of preparing, detecting, and/or assaying an analyte of interest from a sample. The methods utilize functionalized silicon membranes, such as, for example, functionalized silicon nanomembranes. Samples that can be used in the methods may be biological samples, food samples, environmental samples, industrial samples, or a combination thereof. Also provided are kits to perform methods of the present disclosure.

First claim

1 . A method of preparing, detecting, or assaying an analyte of a sample, comprising:
contacting the sample with a fluidic device comprising a functionalized silicon membrane, wherein the fluidic device isolates one or more analyte of interest from the sample; passing a wash solution through the fluidic device; and
i) eluting the isolated analyte of interest;
transferring the eluted analyte of interest to a storage vessel or analytical instrument; and
performing one or more analytical assays on the eluted analyte of interest; or

ii) passing a solution of one or more detection reagent through the fluidic device;
optionally, passing additional wash solution through the fluidic device; and
measuring a signal of one or more detection reagent; or

iii) extracting nucleic acids from the analyte captured by the fluidic device;
performing a sequencing and/or amplification reaction, wherein reagents for such reactions are passed into the fluidic device;
optionally, passing a second wash solution through the fluidic device;
optionally, passing a solution of one or more detection reagent through the device;
measuring a signal of one or more amplification and/or sequencing reaction products.

2 . The method of claim 1 , wherein the functionalized silicon membrane is a functionalized silicon nanomembrane.

3 . The method of claim 1 , wherein the sample comprises a biological sample, a food sample, an environmental sample, an industrial sample, or a combination thereof.

4 . The method of claim 1 , wherein the fluidic device further comprises one or more fluidic channels and/or chambers in fluidic contact with one or more membrane surfaces, one or more aperture having one or more surface, a plurality of nanopores, micropores, or microslits of the membranes.

5 . The method of claim 4 , wherein at least a first and second fluidic channels and/or chambers are in fluidic contact with each other via the one or more aperture and the plurality of nanopores, micropores, or microslits.

6 . The method of claim 5 , wherein the contacting comprises contacting the sample with a first membrane surface and a first fluidic channel or chamber.

7 . The method of claim 5 , wherein the contacting comprises contacting the sample with a second membrane surface, the one or more aperture, and a second fluidic channel or chamber.

8 . The method of claim 1 , wherein any of the steps comprise gravity flow, hydrostatic pressure, pumping, vacuum, centrifugation, gas pressurization, normal flow, tangential flow, or a combination thereof.

9 . The method of claim 1 , wherein washing comprises addition of a buffer solution of specified pH, salt, detergent, and/or carrier biomolecule concentration.

10 . The method of claim 1 , wherein the eluting step comprises chemical denaturation, mechanical denaturation, thermal denaturation, photolysis of a liable bond, reverse flow, or a combination thereof.

11 . The method of claim 1 , wherein adding detection reagent comprises sequential or concurrent addition of one or more solution of biomolecule conjugate, a chromogenic substrate, a chemiluminescent substrate, a co-reagent, or a combination thereof.

12 . The method of claim 1 , wherein adding detection reagent comprises sequential or concurrent addition of at least one or more non-conjugated detection reagents, at least one or more conjugated detection reagents, a chromogenic substrate, a chemiluminescent substrate, a co-reagent, or a combination thereof.

13 . The method of claim 1 , wherein measuring a signal of one or more detection reagent comprises an optical modality for one or more emission, luminescence, and/or absorbance signal at a defined wavelength or range thereof.

14 . The method of claim 1 , wherein performing the sequencing and/or amplification reaction comprises the addition of one or more solutions of buffer, salts, detergents, deoxyribonucleotide triphosphates (dNTPs), enzymes, or a combination thereof.

15 . The method of claim 14 , wherein thermal cycling is performed in the fluidic device.

16 . The method of claim 1 , wherein measuring the signal of one or more amplification and/or sequencing reaction products comprises detection of fluorophore incorporating reaction products, release of fluorophores, fluorophore-bound reaction products, chromophore-bound reaction products, or a combination thereof.

17 . The method of claim 1 , wherein measuring the signal of one or more detection reagents further comprises a plasmic-enhanced optical modality for one or more emission, luminescence, and/or absorbance signal at a defined wavelength or range thereof.

18 . The method of claim 1 , wherein the measuring step comprises using electronic interrogation by one or amperometric or impedimetric methods.

19 . The method of claim 1 , further comprising sequential or concurrent addition of one or more solution of a redox agent, a biomolecule conjugated to a redox agent, or a combination thereof.

20 . The method of claim 1 , further comprising sequential or concurrent addition of one or more solution of detection reagents, wherein the detection reagents are at least one or more non-conjugated detection reagent, at least one or more conjugated detection reagent, a redox agent, or a combination thereof.

21 . The method of claim 1 , wherein the functionalized silicon membrane is functionalized by a method comprises:
contacting the silicon membrane with a chemical oxidation reagent; contacting the silicon membrane with an epihalohydrin; contacting the silicon membrane with a catalyst; and contacting the silicon membrane with one or more biomolecule.

22 . The method of claim 21 , wherein the chemical oxidation reagent comprises a base/acid and a redox reagent.

23 . The method of claim 21 , wherein the epihalohydrin is gaseous epichlorohydrin or gaseous epibromohydrin.

24 . The method of claim 23 , wherein the gaseous epihalohydrin has a vapor pressure of 1.3 to 2666.5 Pa.

25 . The method of claim 21 , wherein the catalyst comprises an acid or base.

26 . The method of claim 21 , further comprising contacting the silicon membrane with a spacer compound prior to contacting the silicon membrane with one or more biomolecules, wherein the spacer compound comprises one or amine group, an aliphatic group having two or more carbons, and one or more additional reactive group.

27 . The method of claim 21 , wherein functionalization of the silicon membrane further comprises:
contacting the silicon membrane with a chemical oxide etchant; contacting the silicon membrane with one or more aldehyde; contacting the silicon membrane with one or more biomolecule; and contacting the silicon membrane with a reductive amination agent.

28 . The method of claim 27 , wherein the chemical oxide etchant comprises a solution of an etchant.

29 . The method of claim 27 , wherein the one or more aldehyde is gaseous and has a vapor pressure of 1.3 to 2666.5 Pa.

30 . The method of claim 27 , wherein the one or more aldehyde comprises a solution having a concentration of 1 μM to 10 M total aldehyde.

31 . The method of claim 28 , further comprising using a dehydration agent.

32 . The method of claim 27 , wherein the reductive amination agent comprises a solution of a reductive agent.

Assignees

• Simpore Inc
• Univ Rochester

Inventors

• Carter Jared A
• Roussie James A
• Madejski Gregory
• Mcgrath James L

Classifications

• B01D71/0215

  Silicon carbide; Silicon nitride; Silicon oxycarbide · CPC title

• B01D67/0093

  Chemical modification · CPC title

• G01N33/54366

  Apparatus specially adapted for solid-phase testing · CPC title

• G01N33/54353

  with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393) · CPC title

• B01D71/82

  characterised by the presence of specified groups, e.g. introduced by chemical after-treatment · CPC title