IPC분류정보
국가/구분 |
United States(US) Patent
등록
|
국제특허분류(IPC7판) |
|
출원번호 |
US-0603413
(2009-10-21)
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등록번호 |
US-8236521
(2012-08-07)
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발명자
/ 주소 |
- Koller, Manfred R.
- Hanania, Elie G.
- Fieck, Anabeth
- Eisfeld, Timothy M.
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출원인 / 주소 |
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대리인 / 주소 |
Knobbe, Martens, Olsen & Bear LLP
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인용정보 |
피인용 횟수 :
0 인용 특허 :
88 |
초록
▼
The invention provides methods for purifying one or more cells based on the level of one or more products secreted by the cells. In one embodiment, the method involves (a) contacting a plurality of cells immobilized in proximity to a capture matrix, the capture matrix capable of localizing a product
The invention provides methods for purifying one or more cells based on the level of one or more products secreted by the cells. In one embodiment, the method involves (a) contacting a plurality of cells immobilized in proximity to a capture matrix, the capture matrix capable of localizing a product secreted by one or more of the cells, with an agent that selectively binds to the product, the agent capable of generating a signal detectable as a property of light; (b) illuminating a population of the cells, the population contained in a frame; (c) detecting two or more properties of light directed from the frame, wherein a first property of light identifies substantially all cells of the population, and the second property of light identifies product localized to the capture matrix; (d) locating (i) substantially all cells of the population with reference to the detected first property of light, and (ii) one or more selected cells with reference to the detected second property of light, and (e) irradiating the non-selected cells, wherein each non-selected cell receives a substantially lethal dose of radiation, whereby one or more selected cells having a desired product secretion profile are purified.
대표청구항
▼
1. A method for purifying cells or groups of cells that secrete a desired product, comprising: culturing a population of cells, wherein the cells are immobilized to or immobilized in the proximity of a capture matrix, such that a detectable amount of product secreted by the immobilized cells remains
1. A method for purifying cells or groups of cells that secrete a desired product, comprising: culturing a population of cells, wherein the cells are immobilized to or immobilized in the proximity of a capture matrix, such that a detectable amount of product secreted by the immobilized cells remains in the vicinity of the cell;labeling said desired product in the vicinity of the cells;imaging the immobilized cells and the label and determining a level of desired product secreted by individual cells or groups of cells; andpurifying selected cells from the population of cells based on the level of desired product secreted by the selected cells. 2. The method of claim 1, wherein the determined level of desired product secreted by individual cells is a relative level. 3. The method of claim 1, wherein determining the level of desired product secreted by individual cells or groups of cells comprises quantitating the level of desired product secreted thereby. 4. The method of claim 1, further comprising storing information for the imaged immobilized cells correlating levels of secretion of desired product with location of the cell or cells producing said levels of secretion. 5. The method of claim 1, wherein the desired product is an antibody. 6. The method of claim 1, wherein the label is fluorescent. 7. The method of claim 1, wherein the desired product is an antibody fragment. 8. The method of claim 1, wherein the desired product is a cytokine. 9. The method of claim 1, wherein the desired product is a growth factor. 10. The method of claim 1, wherein the desired product is an enzyme. 11. The method of claim 1, wherein the desired product is a hormone. 12. The method of claim 1, wherein the desired product is a neurotransmitter. 13. The method of claim 1, wherein the desired product is a protein. 14. The method of claim 1, wherein the desired product is a signaling molecule. 15. The method of claim 1, wherein the desired product is a therapeutic protein. 16. The method of claim 1, wherein the capture matrix comprises one or more of a substance selected from the group consisting of Protein G, Protein A, an antibody, an antibody fragment, an aptamer, and a ligand for the product and a gel. 17. The method of claim 1, wherein the cells are immobilized to or immobilized in the proximity of a capture matrix together with an agent that selectively binds to the desired product, wherein the agent that selectively binds to the desired product is selected from the group consisting of an antibody, an antibody fragment, an aptamer, a substrate, and a ligand. 18. The method of claim 1, wherein the level of desired product is a high level of product relative to other cells of the population. 19. The method of claim 1, wherein the level of desired product is a low level of product relative to other cells of the population. 20. The method of claim 1, wherein the level of desired product is an undetectable level of product.
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