Magnetic separation of magnetized particles from biological fluids

• US 4,910,148 A
• Filed: 12/10/1987
• Issued: 03/20/1990
• Est. Priority Date: 08/21/1987
• Status: Expired due to Term

• Current Assignee: Invitrogen Dynal AS (Invitrogen Dynal Holding AS)
• Original Assignee: Dynal A. S.
• Inventors: Siem, Eivind, Kvalheim, Gunnar, Sorensen, Otto
• Primary Examiner(s): Naff, David M.
• Application Number:
  US07/130,912
• Time in Patent Office: 831 Days
• Field of Search: 210/222, 435/2, 435/7, 435/316, 435/173, 435/239, 435/240.1, 435/240.24, 435/261, 435/284, 435/286, 435/287, 435/803, 435/317.1, 436/526, 436/824, 209/212, 209/213
• US Class Current: 435/317.1
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/0332 using permanent magnets
• 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: C12M 27/16 Vibrating; Shaking; Tilting
• CPC Class Codes: C12M 47/02 Separating microorganisms from the culture medium; Concentration of biomass separating microorganisms from their culture media C12N1/02
• CPC Class Codes: C12N 1/02 Separating microorganisms from their culture media
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

A method and device are provided for separating magnetized particles from biological fluids. Cells such as cancer cells coated with magnetized particles can be separated from uncoated healthy cells. A fluid mixture of cancer cells, healthy cells and magnetizable particles is introduced into a container such as a disposable blood bag which is attached in a cassette on an underlying plane magnetic plane that provides a magnetic field. Incubation is carried out during which the cancer cells become coated with the magnetizable particles. The magnetic field pulls the coated cells down towards the bottom of the bag and anchors them, and uncoated healthy cells are removed from the bag. The separated uncoated healthy cells may be advanced through a final separation unit where any loose magnetizable particles are removed. There is provided means for adjusting vertical distance between the cassette and the magnetic plane and for agitating fluid within the container attached to the cassette. A magnetic screen plate may be placed between the cassette and the magnetic plate to short circuit the magnetic field during incubation. The screen plate is removed to carry out separation.

Claims

1. A device for separating magnetized particles from biological fluids in which said magnetized particles are suspended, said device comprising:

(a) a sample container having an inlet means for introducing biological fluids thereinto and an outlet means for withdrawing biological fluids therefrom,

(b) a magnetic plate means comprising a plurality of low profile, high field density, permanent magnets, wherein the plurality of magnets are oriented on a generally horizontal base plate such that each magnet is spaced from the others and is adjacent magnets having an opposite polarity,

(c) a sample cassette means for holding said sample container in a generally vertical position, said sample cassette means being located above, and generally parallel to, said magnetic plate means,

(d) an adjusting means, operatively connected to said sample cassette means and said magnetic plate means, for adjusting the vertical distance between said sample cassette means and said magnetic plate means, and

(e) a rocker support means for agitating the fluid contents within said sample container.

2. A device in accordance with claim 1, wherein said sample container comprises a sterile, closed bag.

3. A device in accordance with claim 2, wherein said sample container comprises a disposable blood bag.

4. A device in accordance with claim 1, wherein said base plate comprises a magnetizable material.

5. A device in accordance with claim 4, wherein said base plate comprises iron.

6. A device in accordance with claim 1, wherein said plurality of magnets are spaced from one another with spacers, and wherein said spacers are approximately the same vertical thickness as each magnet.

7. A device in accordance with claim 1, wherein the lower portion of said sample cassette means comprises a generally vertical guide means, slidably mounted to said magnetic plate means, for preventing substantial lateral movement of said sample cassette means and for maintaining said sample cassette means in a generally parallel position directly above said magnetic plate means.

8. A device in accordance with claim 1, wherein said adjusting means comprises:

(a) an adjusting frame pivotably mounted to said magnetic plate means,

(b) a tie rod generally parallel to said adjusting frame and pivotably mounted to said magnetic plate means,

(c) a generally horizontal sliding rail pivotably mounted at its opposite ends to said adjusting frame and said tie rod,

and wherein said sample cassette means comprises generally vertical side walls each having a guide dimensioned such that said sliding rail can move horizontally therein.

9. A device in accordance with claim 1, wherein said rocker support means comprises a tiltable table which is in direct contact with the lower portion of said magnetic plate means.

10. A device in accordance with claim 1 further comprising a screen plate, said screen plate being slidably mounted between said sample cassette means and said magnetic plate means, and said screen plate being made of a material which substantially blocks the magnetic field, emanating from said magnetic plate means, from reaching the lower surface of said sample cassette means.

11. A method for separating at least a portion of magnetized particles from biological fluids in which magnetized particles are suspended, said method being carried out with a device which comprises:

a sample container having an inlet means for introducing biological fluids therein and an outlet means for withdrawing biological fluids therefrom;

a magnetic plate means comprising a plurality of low profile, high field density, permanent magnets, wherein the plurality of magnets are oriented on a generally horizontal base plate such that each magnet is spaced from the others and is adjacent magnets having an opposite polarity;

a sample cassette means for holding said sample container in a generally vertical position, said sample cassette means being located above and generally parallel to said magnetic plate means;

an adjusting means, operatively attached to said sample cassette means and said magnetic plate means, for adjusting the vertical distance between said cassette means and said magnetic plate means; and

a rocker support means for agitating the fluid contents within said sample container, said method comprising the steps of;

(a) partially filling said sample container through said inlet means with a biological fluid in which are suspended magnetized particles,

(b) attaching said sample container to said sample cassette means,

(c) adjusting the distance between said sample cassette means and said magnetic plate means with said adjusting means while simultaneously agitating the fluid contents within said sample container with said rocker support means, and

(d) withdrawing said biological fluid through said sample container outlet means while retaining said magnetized particles by said magnetic plate means with said container.

12. A method in accordance with claim 11, wherein said sample container is partially filled with said biological fluid containing magnetized particles after said sample container is affixed to said sample cassette means.

13. A method in accordance with claim 11, wherein a screen plate is positioned between said sample cassette means and said magnetic plate means prior to partially filling said sample container, said screen plate substantially blocking the transmission of a magnetic field from said magnetic plate means to the fluid contents within said sample container.

14. A method in accordance with claim 13, wherein said screen plate is removed from between said sample cassette means and said magnetic plate means after partially filling said sample container with said biological fluids comprising magnetized particles and after said sample container is affixed to said sample cassette means.