Process and installations for separation of magnetic particles in a fluid for biological analysis, and application of said process

• US 6,143,577 A
• Filed: 05/15/1998
• Issued: 11/07/2000
• Est. Priority Date: 05/07/1996
• Status: Expired due to Fees

• Current Assignee: Delta Instruments BV (Perkinelmer Incorporated)
• Original Assignee: Biocom LLC
• Inventors: Bisconte Sconte De Saint Julien, Jean-Claude
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): Do, Pensee T.
• Application Number:
  US08/952,493
• Time in Patent Office: 907 Days
• Field of Search: 436/526, 436/149, 436/806, 436/810, 436/531, 436/534, 436/807, 436/824, 436/528, 435/7.2, 435/7.21, 435/7.23, 435/7.25, 435/7.32, 435/7.34, 435/7.92, 435/7.93, 435/7.94, 435/7.95, 435/971, 210/222, 210/695, 210/223
• US Class Current: 436/526
• CPC Class Codes: B03C 1/035 Open gradient magnetic separators, i.e. separators in which the gap is unobstructed, characterised by the configuration of the gap
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

A process for magnetic immunoseparation of cells, in particular bacteria, fetal cells, stock cells of bone marrow and circulating cancerous cells, of the type consisting in affixing the target cells on paramagnetic balls and in causing a magnetic field to act on a sample containing the affixed cells, the free cells and the surplus paramagnetic balls, in order to isolate the paramagnetic balls, in that the sample is caused to circulate in a tube the section of which is much less than the length over which the magnetic field is applied.

Claims

1. A process for separating cells, comprising:

contacting a sample containing (1) target cells which are affixed to paramagnetic balls, (2) free cells, and (3) surplus paramagnetic balls, with a nonhomogeneous magnetic field,

wherein, during the contacting, the sample is circulated in a tube, wherein the nonhomogeneous magnetic field is applied over a section of the tube measuring less than the length of the tube, and wherein the tube has a proximal portion and a distal portion with respect to fluid flow in the tube,

wherein the surplus paramagnetic balls are immobilized in the proximal portion of the tube and the target cells which are affixed to paramagnetic balls are immobilized in the distal portion of the tube.

2. The process of claim 1, wherein the free cells pass through the tube.

3. The process of claim 1, further comprising contacting cells with paramagnetic balls, wherein the paramagnetic balls are derivatized with a substance capable of binding specifically to a complementary substance on the target cells.

4. The process of claim 1, wherein the target cells are bacteria, fetal cells, stock cells of bone marrow or circulating cancerous cells.

5. The process of claim 1, wherein the sample is circulated in a tube coiled into a spiral in a magnetic-field gradient.

6. The process of claim 1, wherein the section of the tube ranges between 0.5 and 3 millimeters.

7. The process of claim 1, wherein the length of the tube is greater than 10 centimeters.

8. The process of claim 1, wherein the sample is introduced, prior to circulation in the tube, into a reservoir connected to the tube, wherein the reservoir is raised in relation to the tube to provide circulation of the sample by gravity.

9. The process of claim 8, wherein a first volume of sample and a second volume of a rinse liquid are introduced into the reservoir without mixing and without interface layer.

10. The process of claim 1, wherein the preparation is aspirated into the tube placed in the magnetic field gradient, wherein the lower end of the tube is connected to a peristaltic pump.

11. The process of claim 1, wherein magnets which produce the magnetic field are mobile in relation to the tube.

12. The process of claim 1, wherein sample and, optionally, reagents used for the separation, flow in continuous manner in tube at a speed ranging between 0.1 and 10 cm/s.

13. The process of claim 1, further comprising filtering the target cells which are affixed to paramagnetic balls with a microporous membrane filter having a porosity which is greater than the diameter of the paramagnetic balls and less than the diameter of the affixed cells.

14. The process of claim 1, wherein a subsequent filtration stage is undertaken with the aid of a microporous membrane filter the porosity of which is greater than the diameter of the paramagnetic balls and certain surplus cells, and less than the diameter of the affixed cells.