Microscale assays and microfluidic devices for transporter, gradient induced, and binding activities
First Claim
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1. A method of determining an effect of a compound on a transporter activity of a component, the method comprising:
- concomitanly flowing through a first channel a first component comprising the transporter activity, a transportable molecule that interacts with said first component, and a second component comprising a receptor to said transportable molecule that competitively binds with the transportable molecule;
detecting a first signal produced by competitive binding of the receptor to the transportable molecule;
concomitantly flowing through the first channel the first component, the transportable molecule, the second component, and a third component;
detecting a second signal produced by the competitive binding of the receptor to the transportable molecule in the presence of the third component; and
comparing the first and second signals to determine the effects of the third component on the transporter activity of the first component.
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Abstract
Methods of monitoring transporter activity, gradient induced activity, and binding activity in microscale systems, as well as corresponding microscale devices, systems and kits are provided.
248 Citations
19 Claims
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1. A method of determining an effect of a compound on a transporter activity of a component, the method comprising:
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concomitanly flowing through a first channel a first component comprising the transporter activity, a transportable molecule that interacts with said first component, and a second component comprising a receptor to said transportable molecule that competitively binds with the transportable molecule;
detecting a first signal produced by competitive binding of the receptor to the transportable molecule;
concomitantly flowing through the first channel the first component, the transportable molecule, the second component, and a third component;
detecting a second signal produced by the competitive binding of the receptor to the transportable molecule in the presence of the third component; and
comparing the first and second signals to determine the effects of the third component on the transporter activity of the first component. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19)
flowing the transportable molecule into the first channel from a second channel, and flowing the second component into the first channel from a third channel;
wherein the second and third channels intersect the first channel in a mixing region, wherein the first, second and third components diffuse into contact in the mixing region.
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11. The method of claim 1, the method comprising concomitantly flowing the transportable molecule and the second component into the first channel.
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12. The method of claim 1, wherein the first component, the second component, the third component, and the transportable molecule are flowed using one or more fluid direction component comprising one or more of:
- a fluid pressure force modulator, an electrokinetic force modulator, a capillary force modulator and a fluid wicking element.
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13. The method of claim 1, wherein the first channel is a microchannel.
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14. The method of claim 1, wherein the concentration of the transportable molecule is decreased in solution in the first channel as the first component internalizes the transportable molecule.
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15. The method of claim 1, wherein the detectable signal is selected from:
- a cellular activity, a light emission, a radioactive emission, a change in pH and a change in temperature.
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16. The method of claim 1, further comprising varying the concentration of one or more of:
- the first component, the transportable molecule, the third component, and the second component in the first channel and measuring the resulting increase or decrease in signal strength.
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18. The method of claim 1, wherein the third component is a modulator that enhances or inhibits the transporter activity of the first component.
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19. The method of claim 18, the method further comprising flowing the third component into contact with the first component in the first channel prior to introduction of the second component and the transporter molecule.
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17. A kit comprising a first and second component, the first component comprising a transporter activity and the second component being capable of producing a signal upon exposure to a transportable molecule which is transportable by the first component, the components comprising a carrier moiety or set of carrier moieties, the carrier moiety or set of carrier moieties comprising a receptor or transporter, the carrier or carrier set comprising one or more of:
- a cell, a liposome, an organelle, a protein, and a protein-lipid complex.
Specification