Methods for separating particles and/or nucleic acids using isotachophoresis

Patent metadata

Field	Value
Publication number	US-9285340-B2
Application number	US-54221509-A
Country	US
Kind code	B2
Filing date	Aug 17, 2009
Priority date	Aug 17, 2009
Publication date	Mar 15, 2016
Grant date	Mar 15, 2016

Abstract

According to one embodiment, a method includes co-feeding fluids comprising a leading electrolyte, a trailing electrolyte, and at least one of DNA and RNA to a channel, and applying an electric field to the fluids in a direction perpendicular to an axis of the channel for inducing transverse isotachophoresis. In another embodiment, a method includes co-feeding fluids to a channel. The fluids include a leading electrolyte, a trailing electrolyte, biological objects, at least one of DNA and RNA, and a spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of at least some of the biological objects and an electrophoretic mobility of the at least one of the DNA and the RNA. The method also includes applying an electric field to the fluids in a direction perpendicular to an axis of the channel for inducing transverse isotachophoresis. Other methods of isotachophoresis are disclosed in addition to these.

First claim

What is claimed is:

1. A method, comprising:
co-feeding fluids comprising a leading electrolyte, a trailing electrolyte, at least one protein and at least one of DNA and RNA to a channel;
applying an electric field to the fluids in a direction perpendicular to an axis of the channel for inducing transverse isotachophoresis; and
acoustically focusing one or more of the leading electrolyte, the trailing electrolyte, the at least one protein, the DNA, the RNA,
wherein the trailing electrolyte is co-fed to the channel in a mixture with the at least one protein and the at least one of DNA and RNA.

2. The method of claim 1 , wherein either the leading electrolyte is co-fed to the channel in a mixture with a spacer electrolyte; or the trailing electrolyte is co-fed to the channel separately from the at least one of DNA and RNA.

3. The method of claim 1 , further comprising generating a pressure gradient across the fluids as the fluids flow.

4. The method of claim 3 , wherein generating the pressure gradient comprises one or more of:
co-feeding the fluids under an amount of pressure higher than atmospheric pressure applying a vacuum to the channel; and
applying a gravitational force to the fluids in the channel.

5. The method of claim 1 , further comprising applying a vacuum to an outlet of the channel.

6. The method of claim 1 , further comprising co-feeding a spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the leading electrolyte and an electrophoretic mobility of the at least one protein and the at least one of the DNA and the RNA.

7. The method of claim 1 , further comprising co-feeding a spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the trailing electrolyte and an electrophoretic mobility of at least one of the at least one protein and the at least one of the DNA and the RNA, and
wherein the spacer electrolyte is fed to the channel in a mixture with the leading electrolyte.

8. The method of claim 1 , further comprising:
co-feeding a first spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the leading electrolyte and an electrophoretic mobility of the at least one protein and the at least one of the DNA and the RNA; and
co-feeding a second spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the trailing electrolyte and the electrophoretic mobility of the at least one protein and the at least one of the DNA and the RNA.

9. The method of claim 1 , further comprising co-feeding a spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of biological objects in one of the fluids and an electrophoretic mobility of the at least one of the at least one protein and the DNA and the RNA,
wherein the biological objects are selected from a group consisting of viruses, bacteria, and cells.

10. The method of claim 1 , further comprising:
co-feeding a first spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the leading electrolyte and an electrophoretic mobility of the at least one protein and the at least one of the DNA and the RNA;
co-feeding a second spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the trailing electrolyte and the electrophoretic mobility of the at least one protein and the at least one of the DNA and the RNA; and
co-feeding a third spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of at least some biological objects in one of the fluids and the electrophoretic mobility of the at least one protein and the at least one of the DNA and the RNA.

11. The method as recited in claim 10 , further comprising:
separating the leading electrolyte, the trailing electrolyte, the at least one protein, the at least one of the DNA and the RNA, the first spacer electrolyte, the second spacer electrolyte, and the third spacer electrolyte into individual fluid flows within the channel; and
extracting, via a plurality of outlets, each of the leading electrolyte, the trailing electrolyte, the at least one protein, the at least one of the DNA and the RNA, the first spacer electrolyte, the second spacer electrolyte, and the third spacer electrolyte from each of the individual fluid flows within the channel,
wherein the leading electrolyte, the trailing electrolyte, the at least one protein, the at least one of the DNA and the RNA, the first spacer electrolyte, the second spacer electrolyte, and the third spacer electrolyte are co-fed to the channel in a mixture.

12. A method, comprising:
co-feeding fluids to a channel, the fluids comprising:
a leading electrolyte;
a trailing electrolyte;
biological objects;
DNA and RNA, wherein the trailing electrolyte is co-fed to the channel in a mixture with the DNA and the RNA; and
a spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of at least some of the biological objects and an electrophoretic mobility of the DNA and the RNA; and

applying an electric field to the fluids in a direction perpendicular to an axis of the channel for inducing transverse isotachophoresis;
acoustically focusing one or more of the leading electrolyte, the trailing electrolyte, the biological objects, the DNA and the RNA, and the spacer electrolyte;
separating the leading electrolyte, the trailing electrolyte, the biological objects, the DNA and the RNA, and the spacer electrolyte into individual fluid flows within the channel; and
extracting, via a plurality of outlets, each of the leading electrolyte, the trailing electrolyte, the biological objects, the DNA and the RNA, and the spacer electrolyte from each of the individual fluid flows within the channel.

13. The method of claim 12 , wherein the leading electrolyte is co-fed to the channel in a mixture with the spacer electrolyte.

14. The method of claim 12 , wherein the fluids are fed to the channel as a mixture under an amount of pressure higher than atmospheric pressure.

15. The method of claim 12 , wherein the spacer electrolyte is fed to the channel in a mixture with the leading electrolyte.

16. The method of claim 12 , wherein a bulk of the leading electrolyte is spatially separated from the DNA and the RNA upon introduction thereof to the channel.

17. The method of claim 12 , the fluids fu her comprising one or more of:
a first spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the leading electrolyte and the electrophoretic mobility of the DNA and the RNA; and
a second spacer electrolyte having an electrophoretic mobility that is between an electrophoretic mobility of the trailing electrolyte and the electrophoretic mobility of the DNA and the RNA, and
wherein the fluids are fed to the channel as a mixture.

18. The method of claim 12 , further comprising generating a pressure gradient across the fluids as the fluids flow, wherein generating the pressure gradient comprises one or more of:
co-feeding the fluids under an amount of pressure higher than atmospheric pressure;
applying a vacuum to the channel; and
applying a gravitational force to the fluids in the channel.

19. The method of claim 12 , the fluids further comprising;
co-feeding a first spacer electrolyte having electrophoretic mobility that is between an electrophoretic mobility of the leadin

Assignees

• Jung Byoungsok
• Ness Kevin
• Rose Klint A
• L Livermore Nat Security Llc

Inventors

• Jung Byoungsok
• Ness Kevin
• Rose Klint A

Classifications

• G01N27/44773

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

• G01N27/44747 Primary

  Composition of gel or of carrier mixture · CPC title

• G01N2030/8827

  involving nucleic acids · CPC title