Devices and methods for controlling reversible chemical reactions at solid-liquid interfaces by rapid preconcentration and phase replacement

Patent metadata

Field	Value
Publication number	US-9574232-B2
Application number	US-201514630623-A
Country	US
Kind code	B2
Filing date	Feb 24, 2015
Priority date	Feb 25, 2014
Publication date	Feb 21, 2017
Grant date	Feb 21, 2017

Abstract

Devices and methods for controlling reversible chemical reactions at solid-liquid interfaces are disclosed. In particular, the invention relates to a method of increasing reaction rates by concentrating a target molecule in a liquid phase in the region of a reactant or ligand immobilized on a solid followed by removal of the liquid phase and replacement with an immiscible phase, such as an immiscible gas or liquid to impede the reverse reaction. Devices for performing this method to increase the rates and degree of completion of kinetically limited ligand binding or nucleic acid hybridization reactions in affinity chromatography and microarray applications are also disclosed.

First claim

What is claimed is:

1. A method of increasing a rate and a degree of completion of a reaction between a first reactant in a liquid phase and a second reactant on a surface of a solid, the method comprising:
a) contacting the solid with the liquid phase;
b) concentrating the first reactant in the liquid phase near the second reactant on the surface of the solid by performing isotachophoresis (ITP) within the liquid phase using a trailing electrolyte (TE) and a leading electrolyte (LE), such that the first reactant in the liquid phase is concentrated at a LE-TE interface near the second reactant on the surface of the solid;
c) reacting the first reactant in the liquid phase with the second reactant on the surface of the solid; and
d) removing the liquid phase by displacing the liquid phase with an immiscible liquid or a gas, thereby impeding the reverse reaction and increasing the rate and the degree of completion of the reaction.

2. The method of claim 1 , wherein the reaction comprises binding of the first reactant to the second reactant.

3. The method of claim 2 , wherein the first reactant is a target molecule and the second reactant is an affinity ligand immobilized on the solid.

4. A device for performing ITP in combination with affinity chromatography according to the method of claim 3 , the device comprising:
a) a tube comprising an input end and an output end;
b) an affinity region comprising a plurality of affinity ligands immobilized on a solid support, wherein the affinity region is located inside the tube;
c) a first reservoir comprising the TE, wherein the first reservoir is connected to the input end of the tube; and
d) a second reservoir comprising the LE, wherein the second reservoir is connected to the output end of the tube.

5. The device of claim 4 , wherein the solid support comprises a porous polymer monolith (PPM).

6. The device of claim 5 , wherein the PPM comprises an acrylate, methacrylate, vinylpyridine, vinylpyrrolidone, vinylbenzene, divinylbenzene, or vinyl acetate polymer.

7. The device of claim 6 , wherein the PPM comprises poly(glycidyl methacrylate-co-ethylene dimethacrylate) (GMA-EDMA).

8. The device of claim 5 , wherein the PPM has pores of at least 1 micron in diameter.

9. The device of claim 4 , wherein the tube is a capillary, chromatography column, or a microfluidic channel.

10. The device of claim 4 , wherein the affinity region has a hydrodynamic resistance low enough to allow the tube to be filled without the use of a high pressure pump.

11. A method for performing affinity chromatography with the device of claim 4 , the method comprising:
a) filling the second reservoir and the tube with a solution comprising the LE;
b) adding a solution comprising the target molecule and the TE to the first reservoir;
c) performing isotachophoresis (ITP), such that the target molecule is concentrated at a LE-TE interface in the affinity region;
d) capturing the target molecule by binding the target molecule to an affinity ligand in the affinity region;
e) removing the solution comprising the LE and the solution comprising the TE by adding an immiscible liquid or gas to the tube; and
f) eluting the target molecule.

12. The method of claim 2 , wherein the first reactant is a target nucleic acid and the second reactant is a nucleic acid probe immobilized on the solid.

Assignees

• Univ Leland Stanford Junior

Inventors

• Han Crystal
• Santiago Juan G
• Shkolnikov Viktor

Classifications

• G01N27/44795

  Isoelectric focusing · CPC title

• G01N30/60

  Construction of the column · CPC title

• G01N27/44791

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

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• G01N27/27

  Association of two or more measuring systems or cells, each measuring a different parameter, where the measurement results may be either used independently, the systems or cells being physically associated, or combined to produce a value for a further parameter · CPC title