Cell isolation and screening device and method of using same

• US 7,169,577 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, Casagrande, Rocco, Wang, Evelyn
• Primary Examiner(s): Lankford, Jr.; Leon B.
• Assistant Examiner(s): Ford; Allison M.
• Application Number:
  US10/206,340
• Publication Number:
  US 20030032002A1
• 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: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• 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

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 cell(s) to a plurality of wells of a cell expansion device and then detecting the potential desired biological activity of the cell(s). Each of the receptacles comprise a recess sized to isolate a single cell or small group of cells and each of the wells 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 encompassing a recess and bioaffinity ligands, each of the cell isolation regions being sized to isolate about one to about five cells on a surface within said recess, and 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;

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, inverting said cell isolation device, and transferring the about one cell to about five cells from said surface of said cell isolation regions to the wells;

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

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

2. The method of claim 1, wherein the step of delivering comprises delivering one cell to about five cells through a microfluidic channel.

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

4. The method of claim 1, wherein the recess of each of the cell isolation regions defines a top conical portion, and a bottom microwell portion adjoining the top conical portion in a test orientation of the cell isolation device.

5. The method of claim 1, wherein the recess of each of the cell isolation regions defines a top cylindrical portion, an intermediate conical portion, and a bottom microwell portion, the intermediate conical portion being disposed between and adjoining the top cylindrical portion and the bottom microwell portion.

6. The method of claim 1, wherein the bioaffinity ligands are selected from the group consisting of antibodies, self-assembled monolayers (SAMs), lectin, carbohydrate, transmembrane proteins, and antigens.

7. The method of claim 1, wherein the predetermine pitch of the cell isolation regions and the predetermined pitch of the wells matches a pitch of a standard microtiter plate.

8. The method of claim 1, wherein the cells are transferred from the cell isolation device to the cell expansion device by centrifugal force.

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

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

providing a detection device comprising a member defining a plurality of prongs, the prongs being 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.

11. 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 cell isolation region encompassing a recess and bioaffinity ligands, each of the cell isolation regions being sized to isolate about one to about five cells therein, and the cell isolation regions further defining a predetermined pitch with respect to one another; 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, and the wells define a predetermined pitch matching the predetermined pitch of the cell isolation regions wherein when said cell isolation device is placed in direct contact with said cell expansion device and said cell isolation device is inverted said one cell to about five cells may be transferred from said surface of said cell isolation regions to said wells.

12. The kit of claim 11, further comprising a detection device for screening the cells for the desired biological activity.

13. The kit of claim 11, wherein the recess of each of the cell isolation regions defines a top conical portion, and a bottom microwell portion adjoining the top conical portion in a test orientation of the cell isolation device.

14. The kit of claim 13, wherein the top conical portion has a depth of about 1 millimeter.

15. The kit of claim 13, wherein the bottom microwell portion has a diameter of about 10 microns to about 50 microns and a depth of about 10 microns to about 50 microns.

16. The kit of claim 15, wherein the bottom microwell portion has a diameter of about 20 microns and a depth of about 20 microns.

17. The kit of claim 11, wherein the recess of each of the cell isolation regions defines a top cylindrical portion, an intermediate conical portion, and a bottom microwell portion, the intermediate conical portion being disposed between and adjoining the top cylindrical portion and the bottom microwell portion.

18. The kit of claim 17, wherein the cylindrical portion has a diameter of about 2 millimeters and a depth of about 3 millimeters.

19. The kit of claim 17, wherein the conical portion has a depth of about 1 millimeters.

20. The kit of claim 17, wherein the bottom microwell portion has a diameter of about 10 microns to about 50 microns and a depth of about 10 microns to about 50 microns.

21. The kit of claim 11, wherein the pitch of the cell isolation regions and the pitch of the wells match a pitch of a standard microtiter plate.