Apparatus and method for real-time measurement of cellular response
First Claim
1. A method for identifying compounds having biological activity, comprising the steps of:
- (a) combining a homogeneous suspension of living cells with a test compound having an unknown cellular effect to form a test mixture;
(b) directing the test mixture through a detection zone; and
(c) measuring a cellular response of said suspended cells to said test compound as said test mixture is flowing through said detection zone.
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Accused Products
Abstract
An apparatus and method for real-time measurement of a cellular response of a test compound or series of test compounds (303) on a flowing suspension of cells (349), in which a homogeneous suspension of each member of a series of cell types (349) is combined with a concentration of a test compound (303), directed through a detection zone (355), and a cellular response of the living cells is measured in real time as the cells in the test mixture are flowing through the detection zone (355). The apparatus may be used in automated screening of libraries of compounds, and is capable of real-time variation of concentrations of test and standard compounds and generation of dose/response profiles within a short timespan.
271 Citations
81 Claims
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1. A method for identifying compounds having biological activity, comprising the steps of:
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(a) combining a homogeneous suspension of living cells with a test compound having an unknown cellular effect to form a test mixture; (b) directing the test mixture through a detection zone; and (c) measuring a cellular response of said suspended cells to said test compound as said test mixture is flowing through said detection zone. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. An apparatus for automatically measuring the effect of a plurality of test compounds on living cells, comprising:
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a test compound sampler for sequentially providing samples of multiple test compounds; a cell suspension input for providing a suspension of living cells; a mixing zone, coupled to the test compound sampler, for receiving the samples of the test compounds from the test compound sampler, receiving the suspension of living cells from the cell suspension input and mixing each test compound with the suspension of living cells; and a detector, coupled to the mixing zone, for measuring a cellular response of the suspended cells to each test compound. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66)
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67. A method of characterizing the receptors present in a cell comprising the steps of:
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(a) combining a suspension of cells with a test agent known to influence the activity of a particular receptor to form a test mixture; (b) directing said test mixture through a detection zone; (c) measuring a cellular response of said suspension of living cells to said test agent as said test mixture is flowing through said detection zone wherein a response to a test agent indicates that said cell expresses a receptor known to respond to said test agent; and (d) repeating steps (a)-(c) with a series of test agents until the effects of each test agent has been measured. - View Dependent Claims (68, 69, 70, 71)
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72. A method of confirming that a test compound has an effect on the activity of a receptor comprising the steps of:
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contacting a negative control cell type which lacks said receptor with said test agent to form a negative control mixture; directing said negative control mixture through a detection zone; measuring the cellular response of said suspension of living cells to said test compound as said negative control mixture is flowing through said detection zone; contacting cells of the same cell type as the negative control which have been engineered or induced to express said receptor with said test agent to form a test mixture; directing said test mixture through a detection zone; measuring the cellular response of the cells in the test mixture to said test agent, whereby a difference in the measured response of said cells in said test mixture relative to the measured response of said negative control cells indicates that said test agent has an effect on the activity of said receptor. - View Dependent Claims (73, 74, 75)
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76. A method of determining the activity of one or more receptors in a series of cell types comprising:
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(a) combining a suspension of living cells comprising a member of said series of cell types with an agent known to influence the activity of a particular receptor to form a test mixture; (b) directing the test mixture through a detection zone; (c) measuring the cellular response of said suspension of living cells to said test agent; (d) repeating steps (a)-(c) on each member of the series of cell types until the effect of said test agent has been measured in each cell type of said series. - View Dependent Claims (77, 78, 79, 80, 81)
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Specification