IPC분류정보
국가/구분 |
United States(US) Patent
등록
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국제특허분류(IPC7판) |
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출원번호 |
US-0206340
(2002-07-29)
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발명자
/ 주소 |
- Wang,Evelyn
- Casagrande,Rocco
- Kendale,Amar
- Kim,Enoch
- Ostuni,Emanuele
- Schueller,Olivier
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출원인 / 주소 |
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대리인 / 주소 |
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인용정보 |
피인용 횟수 :
5 인용 특허 :
78 |
초록
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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) t
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.
대표청구항
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We claim: 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 isolat
We claim: 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.
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