Method of coupling binding agents to a substrate surface
AssigneeGE Healthcare Bio-Sciences AB
InventorsPär Säfsten, Mattias Tidare
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Abstract
The present invention relates to a method of coupling multiple binding agents to respective areas of a substrate surface by hydrodynamic addressing, using two laminar fluid flows that flow together in the same direction over the substrate surface with an interface to each other to successively couple the binding agents to the substrate areas, wherein each successive coupling of a binding agent to a surface area is followed or preceded by selective deactivation or activation of a selected surface area according to a defined protocol. The invention also relates to the use of such a binding agent-coupled substrate surface for analytical purposes.
Claims20
A method of coupling at least three different binding agents to respective defined areas of a substrate surface that is provided in a Y-shaped flow cell, for subsequent analysis to detect interaction between said binding agents and one or more analytes, by hydrodynamic addressing based on two laminar fluid flows that flow together in the same direction over the substrate surface with an interface to each other, and selectively contacting a defined area of the substrate surface with a desired fluid by positioning the interface laterally through adjustment of the relative flow rates of the two fluid flows, which method comprises coupling at least two of said binding agents to respective defined areas of the substrate surface by at least one of hydrodynamic addressing procedures A and B, wherein:
- procedure A comprises immobilizing a first binding agent to a first area of the substrate surface by contacting the area with a fluid containing the first binding agent, deactivating the first area by subjecting the area to a deactivating fluid, immobilizing a second binding agent to a second area of the substrate surface by contacting a substrate surface area including the first and second areas with the second binding agent, deactivating at least a portion of the first and second areas by subjecting the first and second areas to the deactivating fluid, and immobilizing a third binding agent to a third area of the substrate surface by contacting a substrate surface area including the first, second, and third areas with the third binding agent;
- and
- procedure B comprises deactivating a first area of the substrate surface by subjecting the area to a deactivating fluid, immobilizing a first binding agent to a second area of the substrate surface by contacting the first and second areas with a fluid containing the first binding agent, activating at least a part of the first area by subjecting the area to an activating fluid, immobilizing a second binding agent to the first area by contacting the area with a fluid containing the second binding agent, deactivating at least a portion of the first and second areas by subjecting the first and second areas to the deactivating fluid, activating a portion of a third area of the substrate surface by subjecting the third area to the activating fluid, and immobilizing a third binding agent to a third area of the substrate surface by contacting the third area of the substrate surface with the third binding agent, wherein the first, the second and the third binding agents are different from each other.
Dependencies are inferred from the claim text.