Hydrogel platform for aqueous two-phase concentration of a target to enhance its detection

Patent metadata

Field	Value
Publication number	US-2019187140-A1
Application number	US-201716326687-A
Country	US
Kind code	A1
Filing date	Aug 21, 2017
Priority date	Aug 22, 2016
Publication date	Jun 20, 2019
Grant date	—

Abstract

It was discovered that hydrogel scaffolds can be used to induce phase separation as aqueous two-phase systems (ATPSs) pass through and/or rehydrate the scaffolds, allowing for concentration of target analyte(s) (e.g., biomolecule(s)) into a particular phase of the ATPS or into a leading front. Accordingly, in various embodiments methods and devices are provided that utilize aqueous two-phase systems and hydrogel scaffolds to improve the sensitivity of assays (e.g., of point-of-care assays) without sacrificing cost or ease of use.

First claim

What is claimed is:

1 . A system for the separation and/or concentration of an analyte, said system comprising:
an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase where, in use, said first phase solution becomes a leading phase and said second phase solution becomes a lagging phase; and a hydrogel where said hydrogel and ATPS are disposed to permit said ATPS to flow through said hydrogel.

2 . The system of claim 1 , wherein said system is configured so the hydrogel is substantially dry and is rehydrated as the ATPS flows through said hydrogel.

3 . The system of claim 1 , wherein said hydrogel is hydrated prior to passage of said ATPS through said hydrogel.

4 . The system according to any one of claims 1 - 3 , wherein said hydrogel comprises a naturally-occurring polymer.

5 . The system according to any one of claims 1 - 3 , wherein said hydrogel comprises a synthetic polymer.

6 . The system according to any one of claims 1 - 5 , wherein said hydrogel comprises a hydrogel selected from the group consisting of polylethylene glycol hydrogels, polyethylene oxide hydrogels, polyphosphazene hydrogels, collagen hydrogels, polysaccharide hydrogels, hydroxyethyl methacrylate hydrogels, acrylic hydrogels, copolymers of polyoxyethylene/polyoxypropylene/polyoxyethylene hydrogels, alginate hydrogels, gelatin based hydrogels, chitosan based hydrogels, dextran-aldehyde conjugate hydrogels, hyaluronan/gelatin hydrogels, acrylamide/itaconic acid copolymer hydrogels, acrylic hydrogels, nanometal hydroxide hydrogels, poly(N-vinyl pyrrolidone) hydrogels, poly(N-isopropylacrylamide) hydrogels, collagen-chondroitin sulfate hyaluronic acid hydrogels, polyacrylic acid hydrogels, polyvinyl alcohol hydrogels.

7 . The system according to any one of claims 1 - 3 and 5 , wherein said hydrogel comprises a hydrogel that is a PEGDMA or a PEGUDM hydrogel.

8 . The system of claim 7 , wherein said hydrogel comprises polyethylene glycol dimethacrylate.

9 . The system according to any one of claims 1 - 8 , wherein said hydrogel is a hydrogel formed using a porogen.

10 . The system of claim 9 , wherein said hydrogel is formed using a porogen selected from the group consisting of salt crystals, beads, sodium bicarbonate, sugars, paraffin, and gelatin.

11 . The system of claim 9 , wherein said hydrogel is formed using a porogen that comprises a salt.

12 . The system according to any one of claims 1 - 11 , wherein said ATPS is selected from the group consisting of a polymer/salt ATPS, a polymer/polymer ATPS, a micellar/polymer ATPS, a micellar ATPS, a micellar/salt ATPS, a micellar/sugar ATPS, and an ionic liquid/salt ATPS.

13 . The system of claim 12 , wherein a first phase of solution of said ATPS comprises a Component 1 of Table 1 and a second phase solution of said ATPS comprises a Component 2 of Table 1.

14 . The system of claim 12 , wherein said ATPS is a polymer/salt ATPS.

15 . The system of claim 14 , wherein said ATPS is a PEG/salt ATPS.

16 . The system of claim 15 , wherein said ATPS is a PEG/potassium phosphate ATPS.

17 . The system of claim 12 , wherein said ATPS is a micellar ATPS.

18 . The system of claim 17 , wherein said ATPS is a surfactant/salt ATPS.

19 . The system of claim 18 , wherein said ATPS comprises Triton X-114 (TX-114) surfactant and potassium phosphate salt.

20 . The system of claim 17 , wherein said ATPS comprises a surfactant.

21 . The system of claim 20 , wherein said ATPS comprises TX-114.

22 . The system of claim 12 , wherein said ATPS comprises polypropylene glycol.

23 . The system of claim 12 , wherein said ATPS is a micellar/polymer ATPS.

24 . The system of claim 23 , wherein said ATPS comprises Triton X-100 and dextran.

25 . The system of claim 12 , wherein said ATPS is a micellar/sugar ATPS.

26 . The system of claim 25 , wherein said ATPS comprises Triton X-100 and sucrose.

27 . The system of claim 12 , wherein said ATPS is an ionic liquid ATPS.

28 . The system of claim 27 , wherein said ATPS comprises 1-Butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]).

29 . The system according to any one of claims 1 - 28 , wherein said system is configured to concentrate an analyte in a leading phase of said ATPS.

30 . The system according to any one of claims 1 - 28 , wherein said system is configured to concentrate an analyte in a trailing phase of said ATPS.

31 . The system according to any one of claims 1 - 28 , wherein said system is configured to concentrate an analyte at an interface between a leading phase and a trailing phase of said ATPS.

32 . An assay device for the detection and/or quantification of an analyte in a sample, said device comprising:
a concentration component comprising a hydrogel configured to receive and/or contain an ATPS; and a detection component configured to detect and/or quantify an analyte separated and/or concentrated by said concentration component.

33 . The device of claim 32 , wherein said hydrogel is hydrated prior to contact with an ATPS.

34 . The device of claim 32 , wherein said hydrogel is dried and configured to hydrate on contact with an ATPS.

35 . The device according to any one of claims 32 - 34 , wherein said device comprises a lateral flow assay (LFA) or a flow-through (spot) assay.

36 . The device of claim 35 , wherein said device comprises a lateral flow assay (LFA).

37 . The device of claim 36 , wherein said lateral-flow assay comprises a porous matrix disposed in fluid communication with said hydrogel so that a fluid in said hydrogel can pass into said porous matrix.

38 . The device according to any one of claims 36 - 37 , wherein said concentration component, when in use, comprises a system according to any one of claim 1 - 31 .

39 . The device according to any one of claims 36 - 38 , wherein said lateral-flow assay comprises a probe and/or a development reagent.

40 . The device of claim 39 , wherein said lateral-flow assay comprises probe and a development reagent and is configured so that in use said probe associates said development reagent to enhance a signal.

41 . The device according to any one of claims 37 - 40 , wherein said porous matrix is configured to receive and/or contain an ATPS or components thereof and/or said probe, and/or said development reagent.

42 . The device according to any one of claims 36 - 41 , wherein said LFA comprises a conjugate pad, a test line comprising an antibody that binds said analyte, optionally a control line comprising a secondary antibody, optionally an absorbent pad, and optionally a sample pad.

43 . The device of claim 35 , wherein said device comprises a flow-through (spot) assay.

44 . The device of claim 43 , wherein said device comprises a detection component disposed beneath said concentration component and in fluid communication with said hydrogel so that a fluid in said hydrogel can pass into said detection component.

45 . The device according to any one o

Assignees

• Univ California

Inventors

• Kamei Daniel Takashi
• Wu Benjamin Ming
• Pereira David Yuan
• Wu Chloe Michelle
• Yee Matthew Foosing
• Keefe Joshua Anthony
• Pearce Christina Caroline
• Dailamy Amir Adam
• Wong Vincent K
• Le Nguyen Khoi

Classifications

• G01N33/5436

  with ligand physically entrapped within the solid phase (liposomes G01N33/5432; immunological test elements G01N33/54386) · CPC title

• G01N33/545

  Synthetic resin · CPC title

• G01N2333/183

  Flaviviridae, e.g. pestivirus, mucosal disease virus, bovine viral diarrhoea virus, classical swine fever virus (hog cholera virus) or border disease virus · CPC title

• G01N33/48792

  Data management, e.g. communication with processing unit (for in vivo diagnostics A61B5/0002; transmission systems for measured values G08C) · CPC title

• G01N33/56983

  Viruses · CPC title