Multiple arm apparatus and methods for separation of particles

Patent metadata

Field	Value
Publication number	US-2016096128-A1
Application number	US-201514883234-A
Country	US
Kind code	A1
Filing date	Oct 14, 2015
Priority date	Jan 13, 2012
Publication date	Apr 7, 2016
Grant date	—

Abstract

The invention provides apparatus for separation of particles and methods for using the apparatus. In an embodiment, the apparatus includes three arms extending radially from a central reservoir, each arm being associated with a separation electrode. At least one on the arms includes a separation medium. Using a sequence of driving and mobility-changing voltages, target particles can be separated from closely related particles within a sample. For example, single point mutations can be resolved from a sample containing predominantly wild type nucleic acids.

First claim

1 . An apparatus for separating particles comprising:
at least three electrodes circumferentially surrounding a central reservoir, with a separation medium between at least one electrode and the central reservoir.

2 . The apparatus of claim 1 , wherein the central reservoir comprises a buffer or an additional separation medium.

3 . The apparatus of claim 1 , wherein the separation medium comprises an affinity agent that has a binding affinity for a target particle.

4 . The apparatus of claim 1 , further comprising a controller for regulating the voltage applied to each electrode.

5 . The apparatus of claim 1 , further comprising an arm between each of the at least three electrodes and the central reservoir, at least one of the arms comprising the separation medium.

6 . The apparatus of claim 5 , wherein each arm extends radially outwardly from the central reservoir.

7 . The apparatus of claim 6 , wherein each arm is symmetrically spaced around the central reservoir.

8 . The apparatus of claim 5 , wherein two arms comprise the separation medium.

9 . The apparatus of claim 5 , wherein each arm comprises the separation medium.

10 . The apparatus of claim 5 , wherein the arm comprising the separation medium comprises a loading reservoir adjacent the separation medium.

11 . The apparatus of claim 10 , wherein the loading reservoir extends perpendicularly relative to the length of the separation arm.

12 . The apparatus of claim 11 , further comprising a loading buffer chamber adjacent the separation medium and opposite the loading reservoir.

13 . The apparatus of claim 12 , comprising a first loading electrode in the loading reservoir and a second loading electrode in the loading buffer chamber.

14 . The apparatus of claim 5 , further comprising a buffer chamber at the distal end of each separation arm, each of the at least three electrodes being located in a respective buffer chamber.

15 . A cassette comprising the apparatus of claim 5 , the cassette comprising a top plate and a bottom plate, wherein the arms interpose the top and bottom plates and the separation medium is between the top plate and the bottom plate.

16 . A method for separating target and background particles comprising:
providing a central reservoir and n separation arms containing a separation medium extending outwardly from the reservoir; loading a sample on at least one of the separation arms; applying an electric field that is low in n−1 of the separation arms and high in one of the separation arms; and varying the configuration of the applied electric field a plurality of times so that on average the electric field is high in each one of the n separation arms 1/n of the time and low in each one of the n separation arms (n−1)/n of the time, wherein n is at least 3.

17 . The method of claim 16 , further comprising maintaining a base temperature of the separation medium at a predetermined level.

18 . The method of claim 17 , wherein the base temperature is maintained in the range of 45° C. to 65° C.

19 . The method of claim 16 , comprising allowing Joule heating to increase the temperature in one of the separation arms while the electric field strength in that separation arm is high.

20 . The method of claim 16 , further comprising applying a washing field to the medium.

Assignees

• Univ British Columbia

Inventors

• Marziali Andrea
• Pel Joel

Classifications

• B03C2201/26

  for use in medical or biological applications · CPC title

• B01L2400/0421

  electrophoretic flow · CPC title

• C12N15/101

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

• B01L2300/1805

  Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks · CPC title

• B01D43/00 Primary

  Separating particles from liquids, or liquids from solids, otherwise than by sedimentation or filtration (flotation processes B03D1/00; drying solid materials or objects F26B) · CPC title