Apparatus and method for removing small molecules and ions from low volume biological samples

• US 6,284,117 B1
• Filed: 12/22/1999
• Issued: 09/04/2001
• Est. Priority Date: 12/22/1999
• Status: Expired due to Term

• Current Assignee: Gamida for Life B.V., Nanogen
• Original Assignee: Nanogen, Inc.
• Inventors: Jimenez, Manuel, Sheldon, Ed, Swanson, Paul, Smolko, Daniel, Westin, Lorelei, Mehta, Prashant P., Bloch, Kenneth A, Landis, Geoffrey C.
• Primary Examiner(s): Phasge, Arun S.
• Application Number:
  US09/469,588
• Time in Patent Office: 622 Days
• Field of Search: 204/543, 204/640, 210/321.87, 210/321.89, 210/638, 210/645
• US Class Current: 204/543
• CPC Class Codes: B01D 15/34 Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
• CPC Class Codes: B01D 15/361 Ion-exchange
• CPC Class Codes: B01D 61/28 Apparatus therefor
• CPC Class Codes: B01D 61/422 Electrodialysis
• CPC Class Codes: B01D 61/48 having one or more compartments filled with ion-exchange material , e.g. electrodeionisation
• CPC Class Codes: B01D 63/068 with flexible membrane tubes
• CPC Class Codes: B01J 47/08 subjected to a direct electric current
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: G01N 2001/4016 being a selective membrane, e.g. dialysis or osmosis
• CPC Class Codes: Y02A 20/124 Water desalination
• CPC Class Codes: Y02A 20/131 Reverse-osmosis

Abstract

This invention provides an apparatus and method for desalting a low volume solution for use in connection with an electronically addressable microarray. The apparatus comprises a tubular molecular weight cut-off membrane embedded within a ion exchange resin filled chamber. The apparatus provides a very high surface to volume ratio of membrane pore surface to exchange resin capacity for absorbing charged molecules. The design facilitates the speedy removal of charged molecules from test solutions with the resultant desalted solution having a very low ionic strength suitable for use in the electronic transport of nucleic acids, proteins, and cells.

Claims

1. An apparatus for lowering the ionic strength of a low volume solution used in

the electronic transport of nucleic acids, proteins, and/or cells comprising:

a tubular molecular weight cut-off membrane having a lumen, ion exchange resin, and a chamber surrounding and housing said membrane and resin wherein said membrane has an entrance and an exit port leading through said chamber, said membrane further embedded in and amongst said resin, and said chamber having entrance and exit ports for exchanging a flowable material into and out of said chamber.

2. An apparatus according to claim 1 wherein said lumen of said tubular membrane has a diameter of between 100 μ

m and 3 mm.

3. An apparatus according to claim 1 wherein said tubular membrane has a molecular weight cut-off of between 1 kDa and 500 kDa.

4. An apparatus according to claim 1 wherein said tubular membrane is embedded among said resin as a linear tube.

5. An apparatus according to claim 1 wherein said tubular membrane is embedded among said resin in the form of a coil or spiral.

6. An apparatus according to claim 1 wherein said resin is selected from the group consisting of cation exchange resin, anion exchange resin, resin containing a buffer molecule, resin containing histidine, a granular exchange resin, and a powder exchange resin.

7. An apparatus according to claim 1 wherein said membrane further comprises electrodes for measuring conductivity of a solution passing along the lumen of said membrane.

18. A Method of lowering the ionic strength of a low volume solution comprising:

a. passing said solution into an apparatus of claim 1 or claim 8; and

b. allowing said solution of (a) to remain in said chamber of claim 1 or 8 for between 3 and 15 minutes;

wherein in the case where said solution is passed into an apparatus of claim 8, applying a current across said electrodialysis electrodes while said solution remains in said chamber; and

c. retrieving said solution.

8. An apparatus for lowering the ionic strength of a low volume solution used in

the electronic transport of nucleic acids, proteins, and/or cells comprising:

a tubular molecular weight cut-off membrane having a lumen, ion exchange resin, electrodialysis electrodes, and a chamber surrounding and housing said membrane, electrodialysis electrodes, and resin wherein said membrane has an entrance and an exit port leading through said chamber, said membrane further embedded in and amongst said resin, said chamber having entrance and exit ports for exchanging a flowable material into and out of said chamber, and said electrodes are positioned in a spaced axial alignment and on opposing sides of said chamber with respect to said membrane.

9. An apparatus according to claim 8 wherein said lumen of said tubular membrane has a diameter of between 100 μ

m and 3 mm.

10. An apparatus according to claim 8 wherein said tubular membrane has a molecular weight cut-off of between 1 kDa and 500 kDa.

11. An apparatus according to claim 8 wherein said tubular membrane is embedded among said resin as a linear tube.

12. An apparatus according to claim 8 wherein said tubular membrane is embedded among said resin in the form of a coil or spiral with a resin bed volume of between 0.05 μ

l and 10 ml.

13. An apparatus according to claim 8 wherein said resin is selected from the group consisting of cation exchange resin, anion exchange resin, resin containing a buffer molecule, resin containing histidine, a granular exchange resin, and a powder exchange resin.

14. An apparatus according to claim 8 wherein said membrane further comprises electrodes for measuring conductivity of a solution passing along the lumen of said membrane.

15. An apparatus according to claim 8 wherein said electrodialysis electrodes provide either a direct current or an alternating current.

16. An apparatus according to claim 15 wherein said direct current has a value of between 2 and 10 volts.

17. An apparatus according to claim 15 wherein said alternating current has an ampimeric flux of between 0.1 mA/cm² and 100 mA/cm² and a frequency of between 0.01 and 0.5 Hz.