Apparatus, systems, and methods for capillary electrophoresis

Patent metadata

Field	Value
Publication number	US-11933759-B2
Application number	US-201715707600-A
Country	US
Kind code	B2
Filing date	Sep 18, 2017
Priority date	Sep 27, 2013
Publication date	Mar 19, 2024
Grant date	Mar 19, 2024

Abstract

An apparatus includes a body portion that defines a reservoir and a set of substantially flexible capillaries. The set of substantially flexible capillaries are fixedly coupled to the body portion and in fluid communication with the reservoir. A connector is configured to be coupled to the body portion to be in fluid communication with the reservoir and the set of substantially flexible capillaries. The connector is further configured to be coupled to a vacuum source. The apparatus is arranged such that at least a part of the body portion is electrically conductive. Methods for separating and detecting an analyte from a biological sample with the apparatus are also provided. For example, methods for separating and detecting one or more proteins from a cellular lysate or a purified protein are also provided.

First claim

What is claimed is:

1. An apparatus, comprising:
a capillary, the capillary being vertically oriented such that a second end of the capillary is disposed directly above a first end of the capillary that is configured to be disposed in a sample well;
a body portion defining a reservoir having a first portion that is disposed above the first end of the capillary and a second portion separated from the first portion by a weir, a bottom surface of the reservoir being horizontally oriented when the capillary is vertically oriented, the reservoir configured to be partially filled with a liquid when the bottom surface of the reservoir is oriented horizontally,
an opening in a top surface of the body portion in fluid communication with the reservoir and the second end of the capillary such that a negative pressure applied to a headspace of the reservoir and the capillary via the opening causes ( 1 ) the liquid from the capillary to be drawn into and partially fill the reservoir and ( 2 ) a sample to be drawn from the sample well into the capillary when the first end of the capillary is disposed in the sample well;
an absorbent material disposed in the second portion of the reservoir; and
a contact liquid level sensor protruding into the first portion of the reservoir, the contact liquid level sensor being electrically conductive such that when the liquid within the reservoir contacts the contact liquid level sensor, a signal indicative of a quantity of the liquid within the reservoir and above the capillary is produced.

2. The apparatus of claim 1 , wherein the absorbent material is configured to inhibit leakage from the reservoir.

3. The apparatus of claim 1 , wherein the contact liquid level sensor extends from a cap of the body portion, the cap and the contact liquid level sensor being electrically conductive and configured to transmit an electric potential to the liquid within the reservoir and onto the sample within the capillary such that analytes within the sample within the capillary are electrophoretically separated.

4. The apparatus of claim 1 , further comprising a vacuum source configured to apply the negative pressure to the headspace of the reservoir and the capillary.

5. The apparatus of claim 1 , further comprising:
the sample well, the first end of the capillary disposed within the sample well and having a vertical orientation, at least a portion of the sample well and at least a portion of the body portion being electrically conductive and configured to apply an electric potential across the sample within the capillary to effect electrophoretic separation of analytes within the sample.

6. The apparatus of claim 1 , wherein the capillary and the body portion are collectively configured to be entirely disposed within a housing of a capillary electrophoresis device.

7. An apparatus, comprising:
a capillary;
a reservoir fluidically coupled to the capillary, the reservoir having a first portion and a second portion separated from the first portion by a weir, an opening to the reservoir in fluid communication with the reservoir and the capillary such that a negative pressure applied to a headspace of the reservoir via the opening causes liquid from the capillary to partially fill the reservoir; and
a contact liquid level sensor protruding from a cap into the first portion of the reservoir, the cap and the contact liquid level sensor being electrically conductive such that (1) when the liquid within the reservoir contacts the contact liquid level sensor, a signal indicative of a quantity of the liquid within the reservoir and above the capillary is produced and (2) the cap and the contact liquid level sensor are configured to transmit an electric potential to the liquid within the reservoir and onto a sample within the capillary such that analytes within the sample within the capillary are electrophoretically separated.

8. The apparatus of claim 7 , further comprising a vacuum source configured to draw the liquid through the capillary, the vacuum source operatively coupled to the reservoir via the opening, the vacuum source configured to maintain the liquid in the capillary based on the quantity of the liquid within the reservoir.

9. The apparatus of claim 7 , further comprising:
a sample well, the capillary vertically positioned within the sample well; and
a vacuum source configured to draw the sample vertically from the sample well into the capillary.

10. The apparatus of claim 7 , further comprising:
an absorbent material disposed in the second portion of the reservoir.

11. A method, comprising:
drawing a sample from a sample well through a capillary and into a cartridge reservoir fluidically coupled to the capillary and disposed above the sample well, the capillary being vertically oriented;
receiving, from a contact liquid level sensor that extends into the cartridge reservoir and is constructed of a conductive material, an electrical signal indicative of a fill level of a liquid within the cartridge reservoir;
applying a vacuum pressure to the cartridge reservoir to maintain the sample in a fixed position within the capillary ; and
applying an electric potential across the capillary to cause analytes within the sample to electrophoretically separate while applying the vacuum pressure to cause the sample to maintain the fixed position.

12. The method of claim 11 , further comprising:
calculating a flow rate through the capillary based on the electrical signal and a duration that the sample was drawn through the capillary.

13. The method of claim 11 , wherein the cartridge reservoir includes a first portion separated from a second portion by a weir, the first portion of the cartridge reservoir containing the contact liquid level sensor, the second portion of the cartridge reservoir containing an absorbent material configured to inhibit the liquid from being drawn out of the cartridge reservoir.

14. The method of claim 11 , wherein:
the capillary is disposed within a housing of a capillary electrophoresis device;
the sample is drawn vertically from the sample well disposed within the housing, through the capillary, and into the cartridge reservoir; and
the capillary enters the cartridge reservoir through a bottom portion of the cartridge reservoir.

15. The method of claim 11 , wherein the electric potential is applied across the capillary via the sample well and a conductive portion of the cartridge reservoir from which the contact liquid level sensor extends.

Assignees

• ProteinSimple

Inventors

• Yang Tom W
• Roach David John

Classifications

• G01N27/44782 Primary

  of a plurality of samples · CPC title

• G01N27/44704

  Details; Accessories · CPC title

• C07K1/26

  Electrophoresis · CPC title

• C07K1/285

  multi dimensional electrophoresis · CPC title

• G01N27/44726

  using specific dyes, markers or binding molecules · CPC title