Methods for obtaining liquid from a solid phase

Patent metadata

Field	Value
Publication number	US-9821248-B2
Application number	US-201313834901-A
Country	US
Kind code	B2
Filing date	Mar 15, 2013
Priority date	Mar 16, 2012
Publication date	Nov 21, 2017
Grant date	Nov 21, 2017

Abstract

A method for obtaining a liquid from a porous solid phase is described. The method comprises forming a liquid seal at a first end of a porous solid phase to which a liquid is bound, wherein liquid of the liquid seal is immiscible with the liquid bound to the solid phase, and applying a pressure differential across the porous solid phase to cause the immiscible liquid to move through the porous solid phase towards a second end of the porous solid phase, thereby displacing the liquid bound to the porous solid phase towards the second end and releasing this liquid from the second end. Recovery of liquid from the solid phase using such methods is increased compared with corresponding methods in which no liquid seal is formed. In preferred embodiments, the liquid used to form the liquid seal is a mineral oil. The methods have particular application in nucleic acid extractions which utilize capture of nucleic acid to a solid phase. Kits and apparatus for performing the methods are also described.

First claim

The invention claimed is:

1. A method for obtaining a liquid from a porous solid phase, comprising:
forming a liquid seal at a first end of a porous solid phase to which a liquid is releasably bound, wherein the liquid seal comprises a layer of an immiscible liquid that is immiscible with the liquid that is releasably bound to the solid phase; and
applying a pressure differential across the porous solid phase to cause the layer of immiscible liquid to move through the porous solid phase towards a second end of the porous solid phase, thereby displacing the liquid that is releasably bound to the porous solid phase towards the second end and releasing said releasably bound liquid from the second end.

2. The method of claim 1 , wherein the immiscible liquid is selected from (i) an immiscible liquid that is less dense than the liquid that is releasably bound to the solid phase, (ii) an immiscible liquid that comprises a hydrophobic liquid, and (iii) a hydrophobic liquid that is a mineral oil.

3. The method of claim 1 , wherein the porous solid phase is within a column.

4. The method of claim 1 , wherein the liquid that is releasably bound to the porous solid phase comprises a component of a biological sample, which it is desired to release the liquid that is releasably bound from the solid phase.

5. The method of claim 4 , wherein the component comprises a nucleic acid.

6. The method of claim 5 , wherein the porous solid phase comprises a material that is selected from (i) a material to which the nucleic acid binds at a lower pH and from which nucleic acid is eluted at a higher pH, (ii) an inorganic oxide, and (iii) silica.

7. The method of claim 4 , wherein either or both of (i) the component comprises a protein, and (ii) the porous solid phase comprises an ion-exchange material for protein purification.

8. The method of claim 1 , wherein the liquid that is releasably bound to the solid phase comprises an elution buffer.

9. The method of claim 1 , wherein the pressure differential is applied for a sufficient time that at least some of the immiscible liquid is released from the second end of the porous solid phase.

10. The method of claim 9 , wherein the released releasably bound liquid and the immiscible liquid that is released from the solid phase are collected in an uncapped collection tube.

11. The method of claim 1 , which further comprises heating the porous solid phase.

12. A method for isolation of a biological component, comprising:
(a) contacting (i) a liquid that comprises a biological component, with (ii) a porous solid phase, under conditions and for a time sufficient to releasably bind the biological component to the porous solid phase;
(b) releasing the releasably bound biological component from the porous solid phase into a liquid that is releasably bound to the porous solid phase;
(c) subsequently forming a liquid seal at a first end of the porous solid phase to which the liquid is releasably bound, wherein the liquid seal comprises a layer of an immiscible liquid that is immiscible with the liquid that is releasably bound to the solid phase; and
(d) applying a pressure differential across the porous solid phase to cause the layer of immiscible liquid to move through the porous solid phase towards a second end of the porous solid phase, thereby displacing the liquid that is releasably bound to the porous solid phase towards the second end and releasing said releasably bound liquid from the second end, and thereby isolating the biological component.

13. The method of claim 1 , which is an automated method.

Assignees

• Cambridge Entpr Ltd

Inventors

• Chua Yii Leng

Classifications

• B01D17/0202

  by ab- or adsorption · CPC title

• B01D11/02

  of solids · CPC title

• C12N15/1017

  by filtration, e.g. using filters, frits, membranes · CPC title

• C12N15/101

  by chromatography, e.g. electrophoresis, ion-exchange, reverse phase · CPC title

• B01D15/245

  Adding materials to the effluents · CPC title