Cell isolation and screening device and method of using same

• US 7,169,578 B2
• Filed: 07/29/2002
• Issued: 01/30/2007
• Est. Priority Date: 07/27/2001
• Status: Expired due to Fees

• Current Assignee: Surface Logix, Inc.
• Original Assignee: Surface Logix, Inc.
• Inventors: Kim, Enoch, Kendale, Amar, Ostuni, Emanuele, Schueller, Olivier, Wang, Evelyn, Casagrande, Rocco
• Primary Examiner(s): Lankford, Jr.; Leon B.
• Assistant Examiner(s): Ford; Allison M.
• Application Number:
  US10/206,341
• Publication Number:
  US 20030032071A1
• Time in Patent Office: 1,646 Days
• Field of Search: 435/326, 435/383, 435/287.2, 435/287.6, 435/288.4, 435/305.2, 435/308.1, 435/70.21, 209/17
• US Class Current: 435/30
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00655 the compounds being bound to magnets embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• 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

Abstract

The present invention provides methods and devices for screening a single cell or a small group of cells for a desired biological activity. In particular, the present invention provides for delivering cell(s) to a plurality of cell isolation regions of a cell isolation device, transferring the cell(s) to a plurality of wells of a cell expansion device and detecting the potential desired biological activity of the cell(s). Each of the cell isolation regions comprise a bioaffinity region capable of binding a single cell or a small group of cell(s). This binding of cell(s) may be accomplished through the use of bioaffinity ligands immobilized in the bioaffinity regions of the cell isolation regions. Preferably, the wells of the cell expansion device encompass a cavity that provides sufficient volume for cell proliferation.

Claims

1. A method of screening cells for a desired biological activity comprising:

providing a cell isolation device defining a plurality of cell isolation regions, each cell isolation region being sized for isolating about one to about five cells on a surface of said cell isolation region, each cell isolation region encompassing a bioaffinity region on said surface, the cell isolation regions positioned to have a predetermined pitch with respect to one another;

delivering the about one cell to about five cells to each of the cell isolation regions through a microfluidic channel;

providing a cell expansion device defining a plurality of wells wherein the position of the wells of the cell expansion device corresponds to the predetermined pitch of the cell isolation regions;

placing said cell isolation device in direct contact with said cell expansion device to form a seal such that fluid communication between cells in adjacent cell isolation regions is inhibited, inverting said cell isolation device, and transferring the about one to about five cells from said surface of said cell isolation regions to the wells;

allowing about one cell to about five cells to proliferate and exhibit a desired biological activity; and

assaying the biological activity of the about one to about five cells.

2. The method of claim 1, wherein the microfluidic channel has a diameter of about 100 microns.

3. The method of claim 1, wherein each of the bioaffinity regions comprises bioaffinity ligands immobilized therein.

4. The method of claim 3, wherein the bioaffinity ligands are selected from the group consisting of antibodies, antigens, self-assembled monolayers (SAMs), lectins, carbohydrates, transmembrane proteins, and transmembrane protein receptors.

5. The method of claim 1, wherein the predetermined pitch of the cell isolation regions matches a pitch of a standard microtiter plate.

6. The method of claim 1, wherein each of the cell isolation regions has a diameter of about 10 to about 30 microns.

7. The method of claim 1, wherein transferring comprises transferring about one cell to about five cells from the cell isolation device to the cell expansion device by centrifugal force.

8. The method of claim 1, wherein the cells are hybridoma cells and the biological activity exhibited by the cells is antibody production.

9. The method of claim 1, wherein assaying comprises assaying hybridoma cells for specific antibody production by:

providing a detection device comprising a member defining a plurality of prongs, the prongs coated with specific antigens;

immersing the prongs into the wells to allow potential binding between specific antibodies produced by the hybridoma cells and the antigens;

removing the prongs from the wells;

immersing the prongs into a detection solution;

detecting the presence of specific antibodies.

10. A kit for screening cells for a desired biological activity, the kit comprising:

a cell isolation device defining a plurality of cell isolation regions, each of the cell isolation regions being sized to isolate about one cell to about five cells therein, and each encompassing a bioaffinity region, the cell isolation device further defining a predetermined pitch with respect to one another;

a microfluidic device; and

a cell expansion device defining a plurality of wells wherein the wells of the cell expansion device correspond to respective regions of the cell isolation device such that the structure of said cell isolation device when placed in direct contact with said cell expansion device creates a seal that inhibits fluid communication between cells in adjacent cell isolation regions.

11. The kit of claim 10, further comprising a detection device for screening the about one to about five cells for the desired biological activity.

12. The kit of claim 10, wherein the bioaffinity regions comprise bioaffinity ligands immobilized therein.

13. The kit of claim 12, wherein the bioaffinity ligands are selected from the group consisting of antibodies, antigens, self-assembled monolayers (SAMs), lectins, carbohydrates, transmembrane proteins, and protein receptors.

14. The kit of claim 10, wherein the surface of the cell isolation device has a substantially planar surface and wherein the bioaffinity regions comprise respective discrete areas of the substantially planar surface.

15. The kit of claim 10, wherein each of the cell isolation regions has a diameter of about 10 microns to about 30 microns in diameter.

16. The kit of claim 10, wherein the cell expansion device comprises a plurality of wells comprising lateral sides canted relative to one another.

17. The device of claim 10, wherein the predetermined pitch of the cell isolation regions matches a pitch of a standard microtiter plate.