Apparatus and methods for binding molecules and cells
AssigneeUniversity of Washington
InventorsBuddy D. Ratner, Xuanhong Cheng, Karl Bohringer, Yanbing Wang, Yael Hanein, Ashutosh Shastry
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Abstract
In one aspect, the present invention provides devices for binding cells or molecules, wherein each device includes (a) a body defining a first surface and a second surface that is located opposite to the first surface; (b) a heater disposed upon the first surface; and (c) a temperature-responsive layer disposed upon the second surface. In another aspect, the present invention provides methods for binding molecules or living cells to a temperature-responsive material.
Claims46
A device for binding cells or molecules, wherein the device comprises:
- (a) a body defining a first surface and a second surface that is located opposite to the first surface;
- (b) multiple heaters disposed upon the first surface, wherein some or all of the heaters are independently controllable;
- and
- (c) a temperature-responsive layer disposed upon the second surface, wherein the temperature-responsive layer is deposited by plasma deposition at a first high deposition power and at least one second low deposition power, and comprises a temperature-responsive material having a first layer that crosslinks and adheres to the second surface at the first high deposition power and at least one second layer that forms at the at least one second low deposition power that can exist in a first state that binds molecules or living cells, and can exist in a second state that binds substantially less molecules or living cells than the first state, and wherein the temperature-responsive material is reversibly convertible to the first state from the second state in response to an effective amount of thermal energy.
A method for binding molecules or living cells to a temperature-responsive material, wherein the method comprises the steps of contacting a temperature-responsive material with a population of molecules or a population of living cells, wherein:
- (a) the temperature-responsive material can exist in a first state that binds molecules or living cells, and can exist in a second state that binds substantially less molecules or living cells than the first state, and wherein the temperature-responsive material is reversibly convertible to the first state from the second state in response to an effective amount of thermal energy;
- (b) the temperature-responsive material exists in the first state when the temperature-responsive material is contacted with the population of molecules or living cells, thereby effecting binding of the molecules or living cells to the temperature-responsive material;
- (c) the temperature-responsive material is deposited by plasma deposition at a first high deposition power and at least one second low deposition power, and has a first layer that crosslinks and adheres to a surface at the first high deposition power and at least one second layer that forms at the at least one second low deposition power.
A method for binding more than one type of molecule or more than one type of living cell to a temperature-responsive material, wherein the method comprises the steps of:
- (a) contacting a temperature-responsive material with a first type of molecules or a first type of living cells, wherein the temperature-responsive material is attached to a device body wherein the device body defines a first surface and a second surface that is located opposite the first surface, wherein a first population of heaters and a second population of heaters are disposed upon the first surface, and wherein the temperature-responsive layer is deposited by plasma deposition at a first high deposition power and at least one second low deposition power, and comprises a temperature-responsive material having a first layer that crosslinks and adheres to the second surface at the first high deposition power and at least one second layer that forms at the at least one second low deposition that can exist in a first state that binds molecules or living cells, and can exist in a second state that binds substantially less molecules or living cells than the first state, and wherein the temperature-responsive material is reversibly convertible to the first state from the second state in response to an effective amount of thermal energy wherein the temperature-responsive material forms a temperature-responsive layer on the second surface;
- (b) activating the first population of heaters to heat a first population of temperature-responsive layer portions, located on the second surface opposite the first population of heaters, so that the first type of molecules or the first type of living cells binds to the first population of temperature-responsive layer portions, said activation occurring before or during the contacting of the temperature-responsive material with the first type of molecules or the first type of living cells;
- (c) removing any of the first type of molecules or the first type of living cells that are not bound to the first population of temperature-responsive layer portions;
- (d) contacting the temperature-responsive material with a second type of molecules or a second type of living cells;
- (e) activating the second population of heaters to heat a second population of temperature-responsive layer portions, located on the second surface opposite the second population of heaters, so that the second type of molecules or the second type of living cells binds to the second population of temperature-responsive layer portions, said activation occurring before or during the contacting of the temperature-responsive material with the second type of molecules or the second type of living cells;
- and
- (f) removing any of the second type of molecules or the second type of living cells that are not bound to the second population of temperature-responsive layer portions.
A method for measuring a response of a population of living cells to an agent, wherein the method comprises the steps of contacting a population of living cells with an agent and measuring a response of the living cells to the agent, wherein the living cells are attached to a temperature-responsive material that can exist in a first state that binds living cells, and can exist in a second state that binds substantially less living cells than the first state, wherein the temperature-responsive material is deposited by plasma deposition at a first high deposition power and at least one second low deposition power, and has a first layer that crosslinks and adheres to a surface at the first high deposition power and at least one second layer that forms at the at least one second low deposition power, wherein the temperature-responsive material is reversibly convertible to the first state from the second state in response to an effective amount of thermal energy, and wherein the temperature-responsive material exists in the first state while the living cells are contacted with the agent.
A method for observing the binding of members of a binding pair, wherein the method comprises the steps of contacting a first member of a binding pair with a second member of a binding pair and observing the binding of the first member of the binding pair with the second member of the binding pair, wherein the first member of the binding pair is attached to a temperature-responsive material that can exist in a first state that binds the first member of the binding pair, and can exist in a second state that binds substantially less first member of the binding pair than the first state, wherein the temperature-responsive material is deposited by plasma deposition at a first high deposition power and at least one second low deposition power, and has a first layer that crosslinks and adheres to a surface at the first high deposition power and at least one second layer that forms at the at least one second low deposition power, wherein the temperature-responsive material is reversibly convertible to the first state from the second state in response to an effective amount of thermal energy, and wherein the temperature-responsive material exists in the first state while the first member of the binding pair is contacted with the second member of the binding pair.
Dependencies are inferred from the claim text.