Device for volume coupling in epitachophoresis

Patent metadata

Field	Value
Publication number	US-12596098-B2
Application number	US-202218556327-A
Country	US
Kind code	B2
Filing date	Apr 20, 2022
Priority date	Apr 21, 2021
Publication date	Apr 7, 2026
Grant date	Apr 7, 2026

Abstract

Epitachophoresis (ETP) methods and devices that improve concentrating samples and/or separating components of samples. ETP methods and devices allow for electromigration in two dimensions. Electromigration of a sample may first occur in a first dimension along a single plane. Electromigration may then continue in a second dimension, which may be different from the first dimension. The volume where the electromigration occurs may significantly reduce from the first dimension to the second dimension. This smaller dimension may allow for increased concentration of samples or improved separation of components of a sample.

First claim

What is claimed is:

1 . A method of concentrating a component from a sample, the method comprising:
applying a voltage difference between a first electrode and a second electrode, wherein:
the first electrode is disposed in a first mixture comprising a first electrolyte and the sample,
the second electrode is disposed in a second electrolyte, and
the first electrolyte is different from the second electrolyte;
flowing, using the voltage difference, the component in a first channel in a first direction, wherein:
the first direction is away from the first electrode and to a second channel that intersects the first channel,
the component is focused into a band,
the band while in the first channel is characterized as having a first thickness perpendicular to the first direction;
flowing, in the second channel, the component focused in the band in a second direction, wherein:
the second electrolyte is in the second channel,
the second channel is an annular space formed between a cavity and a vessel,
the second direction is from the intersection with the first channel to an orifice of the vessel,
the second channel characterized as having a second thickness that is perpendicular to the second direction and extends from a surface of the cavity to an opposing surface of the vessel,
a second mixture comprising the component and the second electrolyte in the second channel, and
the first thickness is greater than the second thickness; and
collecting the second mixture in the vessel while applying the voltage difference, wherein the concentration of the component in the second mixture in the vessel is higher than the concentration of the component in the sample.

2 . The method of claim 1 , wherein:
the component is a first component, the sample comprises a second component, the band is a first band, and the second component is focused into a second band in the first channel, the method further comprising: flowing, using the voltage difference, the second component in the first channel, and flowing, in the second channel, the second component focused into the second band in the second direction.

3 . The method of claim 2 , further comprising collecting the second component in the vessel.

4 . The method of claim 1 , wherein the first thickness is a height.

5 . The method of claim 1 , wherein the annular space is defined by an outer surface of the vessel.

6 . The method of claim 1 , wherein the first channel and the second channel are each defined by a base.

7 . The method of claim 1 , wherein the first direction is not collinear with the second direction.

8 . The method of claim 1 , wherein the vessel comprises a conical shape.

9 . The method of claim 1 , wherein the second electrolyte is disposed in a gel in the first channel.

10 . The method of claim 1 , wherein the band is ring-shaped.

11 . The method of claim 1 , wherein:
the first channel is characterized by a first volume, the second channel is characterized by a second volume, and the first volume is at least 10 times greater than the second volume.

12 . The method of claim 1 , wherein the second channel has a length in the second direction of at least 2 mm.

13 . The method of claim 1 , wherein collecting the second mixture in the vessel comprises flowing the second mixture into the vessel in a third direction against gravity.

14 . The method of claim 1 , wherein flowing, in the second channel, the component focused in the band in the second direction comprises reducing the radius of the band and flowing the component in the direction of gravity.

15 . A system for concentrating components in a sample, the system comprising:
a base defining a first channel and a cavity, wherein the cavity comprises a conical shape with a first outer diameter at a top of the cavity being greater than a second outer diameter at a bottom of the cavity; a first electrode disposed in the first channel; and a second electrode; wherein:
the first channel is in fluid communication with the cavity,
an outer diameter of the first channel is greater than the first outer diameter at the top of the cavity,
the second electrode is configured to be in closer electrical communication with the cavity than with the first channel when the first channel and the cavity contain an electrolyte,
the first channel is characterized by a first volume,
the cavity is configured to receive a vessel and to form a second channel when the vessel is disposed in the cavity,
the second channel is an annular space,
the annular space is defined by a surface of the base and a surface of the vessel when the vessel is disposed in the cavity,
the second channel is characterized by a second volume, and
the first volume is greater than the second volume.

16 . The system of claim 15 , wherein the surface of the base and the surface of the vessel are concentric.

17 . The system of claim 15 , wherein the surface of the base is conical.

18 . The system of claim 15 , wherein the cavity is below a bottom surface of the base defining the first channel.

19 . The system of claim 15 , further comprising a membrane between the cavity and the second electrode.

20 . The system of claim 15 , wherein:
the base defines a plurality of structures extending into the cavity, the plurality of structures includes a first structure and a second structure on the opposite side of the cavity as the first structure, and the distance between the first structure and the second structure is a diameter of the vessel.

Assignees

• Roche Sequencing Solutions Inc

Inventors

• Foret Frantisek
• Voracova Ivona
• Prikryl Jan
• Novotny Jakub

Classifications

• C12Q1/6806

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

• G01N27/44739

  Collecting the separated zones, e.g. blotting to a membrane or punching of gel spots · CPC title

• G01N27/44773 Primary

  Multi-stage electrophoresis, e.g. two-dimensional electrophoresis · CPC title