Multi-mode separation for target detection and cell growth monitoring

• US 10,724,930 B2
• Filed: 10/24/2017
• Issued: 07/28/2020
• Est. Priority Date: 05/03/2012
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Michigan (University of Michigan)
• Original Assignee: Board of Regents of the University of Michigan (University of Michigan)
• Inventors: McNaughton, Brandon H., Kinnunen, Paivo, Younger, John G.
• Primary Examiner(s): Chin, Christopher L
• Application Number:
  US15/792,718
• Publication Number:
  US 20180143112A1
• Time in Patent Office: 1,008 Days
• Field of Search: 422 73, 422 8205, 4352887, 436526, 436536, 436538, 436806, 73 6142, 73 6151
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 for use in medical applications
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

Sandwich separation is based on forming a sandwich complex with a magnetic bead, buoyant bead, and a target. Once a sandwich formation is created, the sandwich can be separated using its dual physical properties, namely magnetism and buoyancy. Sandwich separation is highly specific, allows for removal of the beads that do not have any attached target, and reduces the number of background beads. Sandwich separation can also be used to allow for target detection in raw specimen. Also, improvement of detection capability is accomplished by performing AMBR measurements on a solid interface, where the rotational period speeds up and allows for dramatically reduced time-to-result.

Claims

1. An apparatus comprising:

an inlet;

a separation chamber device coupled to receive a sample from the inlet at a sample flow rate, the sample within the separation chamber containing targets, magnetic beads having a moiety that can specifically associate with the targets under appropriate conditions, buoyant beads having a moiety that can specifically associate with the targets under appropriate conditions, and formed complexes comprising the target and one or more magnetic beads and one or more buoyant beads, the separation chamber having at least one outlet positioned to receive and isolate the formed complexes from a non-complex within the sample; and

a magnet movable relative to a portion of the separation chamber, movable from an adjacent to proximal position relative to the outlet, for affecting specificity of isolation of the formed complexes, the movement of the magnetic resulting in an adjustable magnetic force controllable to isolate the formed complexes based on the magnetic moment of the one or more magnetic beads associated with the targets and based on the buoyancy of the one or more buoyant beads associated with the targets.

2. The apparatus of claim 1, wherein the separation chamber device comprises a plurality of separation chambers, in a sequential configuration, where each successive separation chamber provides further isolation of the formed complexes.

3. The apparatus of claim 1, wherein the separation chamber device is a vial.

4. The apparatus of claim 1, wherein the separation chamber device is rotatable.

5. The apparatus of claim 1, wherein the separation chamber device is integrated with the inlet and outlet.

6. The apparatus of claim 1, wherein the magnet is movable along a length of the separation chamber to adjust the position and isolation of the formed complexes.

7. The apparatus of claim 1, wherein the magnet is a rotatable around the separation chamber.

8. The apparatus of claim 1, wherein the magnet produces a rotating magnetic field.

9. The apparatus of claim 1, further comprising a visualization region for analyzing the formed complexes isolated within the separation chamber.

10. The apparatus of claim 9, wherein the visualization region is positioned to allow illumination of the separation chamber using an external illumination source and positioned to allow detection of resulting emissions from the formed complexes using an external detector.

11. The apparatus of claim 9, wherein the visualization region is positioned to allow illumination of the separation chamber using an integrated illumination source and positioned to allow detection of resulting emissions from the formed complexes using an integrated detector.