Claims
- 1. A method for determining the population of each of a plurality of membrane-associated analytes of a cell, the method comprising the steps of:
providing a column of buffer solution moving along an electrophoresis channel, the channel having in succession a cleavage zone and a separation zone and having an electrical field collinear with the moving column of buffer solution; inserting a cell into the moving column of buffer solution upstream of the cleavage zone so that the cell is transported by the moving column of buffer solution through the electrophoresis channel to the cleavage zone, wherein the cell comprises, attached to each of the plurality of membrane-associated analytes, a binding composition specific for that membrane-associated analyte and having one or more molecular tags, each attached to the binding composition by a cleavable linkage, wherein the molecular tags of different binding compositions have different electrophoretic mobilities, so that distinct peaks are formed upon electrophoretic separation of a mixture of the tags; cleaving the cleavable linkages in the cleavage zone to release the molecular tags from the cell; and electrophoretically separating and identifying the released molecular tags in the moving column of buffer solution in the separation zone of the electrophoresis channel, to determine the populations of membrane-associated analytes of the cell.
- 2. The method of claim 1, wherein a sequence of cells is inserted into the moving column of buffer solution upstream of the cleavage zone, at a frequency such that only one cell is present in the cleavage zone at any given time.
- 3. The method of claim 2, where electrophoretically separating the released molecular tags produces an electropherogram having a series of discrete groups of signals from said released molecular tags, each group corresponding to a single cell.
- 4. The method of claim 1, wherein said cell is inserted into the moving column of buffer solution via a side channel which intersects the electrophoresis channel upstream of the cleavage zone, said side channel being in fluid communication with a sample reservoir.
- 5. The method of claim 4, wherein said electrophoresis channel is in fluid communication with a buffer reservoir, located upstream of said side channel.
- 6. The method of claim 6, wherein pressure is applied simultaneously to said sample reservoir and said buffer reservoir.
- 7. The method of claim 1, wherein said cleavable linkage is photochemically cleavable.
- 8. The method of claim 1, wherein said cell comprising said binding compounds is prepared by contacting a cell with a plurality of binding compounds, each having a binding moiety specific for a different membrane-associated analyte, wherein said cell includes at least one of said membrane-associated analytes.
- 9. The method according to any of claims 1 through 8 wherein said cell is a human cell.
- 10. The method of claim 9 wherein at least one of said membrane-associated analytes is a cell membrane receptor.
- 11. The method of claim 10 wherein said cell membrane receptor is an ErbB receptor.
- 12. The method of claim 9 wherein at least one of said membrane-associated analytes is a receptor dimer.
- 13. The method of claim 12 wherein said receptor dimer is a Her dimer.
- 14. A microfluidics system for determining the population of each of a plurality of membrane-associated analytes of a cell, comprising:
an electrophoresis channel containing a moving column of buffer solution, the electrophoresis channel having in succession a cleavage zone, a separation zone, and a detection zone, and having an electrical field collinear with the moving column of buffer solution; a sample-injection channel for inserting a cell into the moving column of buffer solution upstream of the cleavage zone so that the cell is transported by the moving column of buffer solution through the electrophoresis channel to the cleavage zone; wherein the cell comprises, attached to each of the plurality of membrane-associated analytes, a binding compound specific for the membrane-associated analyte and having one or more molecular tags, each attached to the binding compound by a cleavable linkage, wherein the molecular tags of different binding compounds have different electrophoretic mobilities, so that distinct peaks are formed upon electrophoretic separation of a mixture of molecular tags; a light source having a light beam directed to the cleavage zone for directly or indirectly cleaving the cleavable linkages in the cleavage zone to release the molecular tags from the cell; and a detector for collecting a signal generated by electrophoretically separated molecular tags in the detection zone to determine the populations membrane-associated analytes of the cell.
- 15. The system of claim 14 wherein said cell is a human cell.
- 16. The method of claim 15 wherein at least one of said membrane-associated analytes is a cell membrane receptor.
- 17. The method of claim 16 wherein said cell membrane receptor is selected from the group consisting of receptor tyrosine kinases and G-protein coupled receptors.
- 18. The method of claim 17 wherein said cell membrane receptor is an ErbB receptor.
- 19. The method of claim 15 wherein at least one of said membrane-associated analytes is a receptor dimer.
- 20. The method of claim 12 wherein said receptor dimer is a Her dimer.
Parent Case Info
[0001] This application claims priority from U.S. provisional application Ser. No. 60/425,129 filed 8 Nov. 2002.
Provisional Applications (1)
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Number |
Date |
Country |
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60425129 |
Nov 2002 |
US |