Immunomagnetic assay system for clinical diagnosis and other purposes

• US 5,972,721 A
• Filed: 03/14/1997
• Issued: 10/26/1999
• Est. Priority Date: 03/14/1996
• Status: Expired due to Fees

• Current Assignee: The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: United States Of America As Represented By The Secretary Of The Air Force
• Inventors: Bruno, John G., Kilian, John P., Kiel, Johnathan L.
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/816,429
• Time in Patent Office: 956 Days
• Field of Search: 209/2, 209/3, 209/214, 209/232, 210/222, 210/695, 422/57, 422/58, 422/59, 422/61, 422/68.1, 435/7.2, 435/7.21, 435/7.24, 435/7.32, 436/526, 436/534, 436/63, 436/824
• US Class Current: 436/526
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/032 Matrix cleaning systems
• CPC Class Codes: B03C 1/0332 using permanent magnets
• CPC Class Codes: B03C 1/034 characterised by the matrix elements
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

An apparatus and method for immunomagnetic separation and concentration of target biological materials is disclosed. The immunomagnetic separation is performed by a magnetic flow cell, or filter block, as part of an automated mostly continuous immunomagnetic assay system. The magnetic flow cell has two bundles of ferromagnetic rods or pins positioned inside an internal chamber so that a fluid sample flowing through the flow cell passes through the pins. A pair of cobalt magnets flank the flow cell so that the pins concentrate and sufficiently increase the magnetic fields so that even nanometer size magnetic beads can be captured. The overall system combines a reaction subsystem for reacting coated magnetic beads with a sample, a collection subsystem for capturing magnetic beads, a rinsing subsystem for removing debris and a filtering subsystem for removing captured magnetic beads from the collection subsystem. The new magnetic flow filter is the key component for the collection and filtering subsystems.

Claims

1. A magnetic flow cell for capturing antibody-coated magnetic beads from a fluid sample as part of an immunomagnetic assay system, comprising:

(a) a housing;

(b) a chamber inside the housing;

(c) an inlet port through the housing into the chamber for flowing the fluid sample into the chamber;

(d) an outlet port through the housing out from the chamber for flowing the fluid sample out of the chamber;

(e) a plurality of paramagnetic rods positioned inside the chamber such that the flowing fluid sample will flow past the plurality of the paramagnetic rods as it flows through the chamber,

wherein the chamber is shaped so that its cross-sectional area expands from where it connects to the inlet port to a position past at least the plurality of the paramagnetic rods.

2. The magnetic flow cell according to claim 1, wherein the plurality of the paramagnetic rods are positioned perpendicularly to an axis drawn between the inlet port and the outlet port.

3. An automated immunomagnetic assay system for analysis of an analyte in a sample, comprising:

(a) a reaction subsystem for reacting magnetic beads coated with an antibody, which binds to the analyte, with the sample in a solution to make a fluid sample;

(b) a collection subsystem for capturing the magnetic beads out of the fluid sample;

(c) a rinsing subsystem for removing non-magnetic bead debris from the collection subsystem; and

,

(d) a filtering subsystem for removing the captured magnetic beads from the collection subsystem and holding them for the analysis.

4. The automated immunomagnetic assay system according to claim 3, wherein the collection and filtering subsystems comprise:

(a) a magnetic flow cell, the magnetic flow cell comprising

(i) a housing;

(ii) a chamber inside the housing;

(iii) an inlet port through the housing into the chamber for flowing the fluid sample into the chamber;

(iv) an outlet port through the housing out of the chamber for flowing the fluid sample out of the chamber; and

(v) a plurality of paramagnetic rods positioned inside the chamber perpendicular to an axis drawn between the inlet port and the outlet port such that the flowing fluid sample will flow past the plurality of the paramagnetic rods as it flows through the chamber;

(vi) wherein the chamber is shaped so that its cross-sectional area expands where it connects to the inlet port to a position past at least the plurality of the paramagnetic rods;

and,

(b) a pair of movable magnets positioned such that they are movable to and from a position flanking the magnetic flow cell.

5. An automated immunomagnetic assay system for analysis of an analyte in a sample, comprising:

(a) a reaction subsystem for reacting magnetic beads coated with an antibody, which binds to the analyte, with the sample in a solution to make a fluid sample;

(b) a collection subsystem for capturing the magnetic beads out of the fluid sample;

(c) a rinsing subsystem for removing non-magnetic bead debris from the collection subsystem; and

(d) a filtering subsystem for removing the captured magnetic beads from the collection subsystem and holding them for the analysis;

wherein the collection and filtering subsystems comprise;

(i) a magnetic flow cell, the magnetic flow cell comprising

(1) a housing;

(2) a chamber inside the housing;

(3) an inlet port through the housing into the chamber for flowing the fluid sample into the chamber;

(4) an outlet port through the housing out of the chamber for flowing the fluid sample out of the chamber; and

(5) a plurality of paramagnetic rods positioned inside the chamber such that the flowing fluid sample will flow past the plurality of the paramagnetic rods as it flows through the chamber;

wherein the chamber is shaped so that its cross-sectional area expands from where it connects to the inlet port to a position past at least the plurality of the paramagnetic rods; and

,

(ii) a pair of movable magnets positioned such that they are movable to and from a position flanking the magnetic flow cell.

6. The automated immunomagnetic assay system according to claim 5, wherein the plurality of the paramagnetic rods are positioned perpendicularly to an axis drawn between the inlet port and the outlet port.

7. A method for capturing antibody-coated magnetic beads from a fluid sample as part of an immunomagnetic assay system, comprising the steps of:

(a) flowing the fluid sample through a chamber, wherein the chamber comprises a plurality of paramagnetic rods positioned inside the chamber such that the flowing fluid sample flows past the plurality of the paramagnetic rods as it flows through the chamber and wherein the chamber is shaped so that its cross-sectional area expands from the fluid sample enters the chamber to a position past at least the plurality of the paramagnetic rods; and

,

(b) flanking the chamber with a pair of magnets as the fluid sample flows through the chamber thereby collecting the magnetic beads on the plurality of the paramagnetic rods;

(c) rinsing non-magnetic bead debris from the chamber by flowing a buffer solution through the chamber;

(d) removing the pair of magnets to release the magnetic particles from the plurality of the paramagnetic rods; and

(e) capturing the released magnetic particles on a membrane filter provided in the chamber by reversing the flow of the buffer solution through the chamber.

8. The method for capturing antibody-coated magnetic beads according to claim 7, wherein the plurality of the paramagnetic rods are positioned perpendicularly to the flow of the fluid sample through the chamber.

9. A method for performing an immunomagnetic assay for an analyte in a sample, comprising the steps of:

(a) reacting magnetic beads coated with an antibody, which binds to the analyte, with the sample in a solution to make a fluid sample;

(b) collecting the magnetic beads out of the fluid sample by the steps of;

(i) flowing the fluid sample through a chamber, wherein the chamber comprises a collection subsystem comprising a plurality of paramagnetic rods positioned inside the chamber such that the flowing fluid sample flows past the plurality of the paramagnetic rods as it flows through the chamber and wherein the chamber is shaped so that its cross-sectional area expands from the fluid sample enters the chamber to a position past at least the plurality of the paramagnetic rods; and

,

(ii) flanking the chamber with a pair of magnets as the fluid sample flows through the chamber thereby collecting the magnetic beads on the plurality of the paramagnetic rods;

(c) rinsing non-magnetic bead debris from the collection subsystem by flowing a buffer solution through the chamber;

(d) capturing the collected magnetic beads on a membrane filter provided in the chamber by moving the pair of magnets away from the position flanking the chamber and reverse flowing the buffer solution through the chamber; and

(e) analyzing the analyte bound to the magnetic beads captured on the membrane filter.

10. The method for performing an immunomagnetic assay according to claim 9, wherein the plurality of the paramagnetic rods are positioned perpendicularly to the flow of the fluid sample through the chamber.