Continuous cell detection by isotachophoresis

Patent metadata

Field	Value
Publication number	US-9885688-B2
Application number	US-201414555793-A
Country	US
Kind code	B2
Filing date	Nov 28, 2014
Priority date	Nov 29, 2013
Publication date	Feb 6, 2018
Grant date	Feb 6, 2018

Abstract

The present invention provides a system including: a protein having a domain that binds a membranal component; an inlet for sample flow, an Isotachophoresis (ITP) system and a flow generating means connected or coupled to the aqueous parts of the ITP. The invention also provides a method for detecting and or sorting cells with this system.

First claim

What is claimed is:

1. A system comprising:
(A) a protein having a domain that binds a membranal component;
(B) a first flow channel configured to provide flow of a liquid sample to an isotachophoresis (ITP) apparatus, said liquid sample comprising or suspected of comprising a cell, a cell membrane or a fraction of a cell membrane; and
(C) wherein said ITP apparatus comprises:
(a) a first zone and a second zone, said first zone is configured to contain a solution of high effective mobility leading electrolyte (LE) ion, and said second zone is configured to contain a solution of low effective mobility trailing electrolyte (TE) ion, said first zone and said second zone are configured to be operably connected to at least one anode and at least one cathode; and
(b) at least one second flow channel elongated between said first zone and second zone;

(D) a flow regulator, said flow regulator is configured to generate a flow countering an electromigration of said protein in the first flow channel so as to maintain said protein in a pre-determined zone.

2. The system of claim 1 , wherein said membranal component is a protein.

3. The system of claim 1 , wherein said liquid sample is continuously injected into said first flow channel.

4. The system of claim 1 , wherein said membranal component is a net charge of phospholipid head groups or charge density of phospholipid head groups.

5. The system of claim 1 , wherein said flow countering an electromigration is electroosmotic or pressure driven.

6. The system of claim 1 , wherein said protein is labeled.

7. The system of claim 6 , wherein said protein is labeled by at least one label selected from the group consisting of: fluorescently labeled, chemiluminescently labeled, radioactively labeled, and colorimetrically labeled.

8. The system of claim 1 , wherein said protein is an anti-microbial peptide (AMP).

9. The system of claim 1 , wherein said protein is an antibody or an antibody fragment.

10. The system of claim 1 , comprising an anionic or cationic ITP focusing stationary zone characterized by high concentration of said protein.

11. The system of claim 1 , wherein said LE comprises sodium hydroxide and/or wherein said TE comprises Pyridine.

12. The system of claim 1 , further comprising a collector configured for collecting labeled cells from said second zone.

13. The system of claim 1 , further comprising a detector selected from the group consisting of: a photodetector, a photomultiplier tube (PMT), a conductivity detector, a radioactive detector, a camera and any combination thereof.

14. The system of claim 1 , wherein said cell is an intact cell.

15. A method for detecting a cell of interest, the method comprising the steps of:
a) providing a labeled protein having a domain that binds a membranal component in said cell of interest, wherein said labeled protein is focused by ITP to an ITP zone in a liquid flow channel;
b) applying a counter flow so as to maintain said labeled protein in a stationary zone;
c) providing a sample to said flow channel, said sample comprising or suspected of comprising the cell of interest, a cell membrane or a fraction of a cell membrane thereof; and
d) detecting said cell of interest bound to said labeled protein.

16. The method of claim 15 , wherein said detecting includes labeling, separating, enriching, isolating, or any combination thereof.

17. The method of claim 15 , wherein said detecting is achieved by using a photodetector, a photomultiplier tube (PMT), a conductivity detector, a radioactive detector, a camera or any combination thereof.

18. The method of claim 15 , wherein said counter flow is electroosmotic flow or pressure driven flow.

19. The method of claim 15 , wherein said labeled is:
chemiluminescently labeled, radioactively labeled, or colorimetrically labeled.

20. The method of claim 15 , wherein said labeled protein is an anti-microbial peptide (AMP).

21. The method of claim 15 , wherein said labeled protein is an antibody.

Assignees

• Technion Res & Dev Foundation

Inventors

• Bercovici Moran
• Schwartz Ortal

Classifications

• G01N27/44791

  Microapparatus (sample containers with integrated microfluidic structures B01L3/5027) · CPC title

• B01L2400/0421

  electrophoretic flow · CPC title

• B01L2300/0867

  Multiple inlets and one sample wells, e.g. mixing, dilution · CPC title

• G01N33/561

  Immunoelectrophoresis · CPC title

• B01L2300/0627

  Sensor or part of a sensor is integrated · CPC title