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US 8,507,208 B2Granted
Methods and compositions for detection of biological materials using microfluidic devices
AssigneeUniversity of Wyoming
InventorsRobert C. Corcoran, Debashis Dutta
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Abstract
Provided herein are microfluidic devices and methods useful for sensitive detection of analytes. The methods and devices described herein are also useful for detecting direct or indirect binding of enzymes or catalysts to a surface, for example a surface having analytes bound thereon. Methods disclosed herein include embodiments utilizing a pre-concentration scheme to improve signal levels of corresponding reporter moieties.
Claims21
A method for detecting a target analyte, the method comprising the steps of:
- providing a microfluidic device comprising a binding surface in fluid communication with a microfluidic trapping region and at least two electrodes, wherein the microfluidic trapping region comprises a semi-permeable membrane and wherein the binding surface comprises binding molecules capable of selectively binding to the target analyte;
- providing to the binding surface a first solution comprising the target analyte molecules, wherein at least a number of the target analyte molecules selectively bind to the binding molecules of the binding surface to provide a binding surface having bound target analyte molecules;
- providing to the binding surface having the bound target analyte molecules a second solution comprising catalyst molecules, wherein at least a number of the catalyst molecules further binds directly or indirectly to the target analyte molecules bound to the binding surface;
- providing to the binding surface having the bound target analyte molecules and the catalyst molecules a third solution comprising substrate molecules, wherein at least a number of the substrate molecules undergo a catalytic chemical reaction with the catalyst molecules bound directly or indirectly to the target analyte molecules bound to the binding surface, thereby directly or indirectly producing reaction product molecules having an ionic charge different from an ionic charge of the substrate molecules;
- transporting at least a number of the reaction product molecules into the microfluidic trapping region by applying an electrical potential between the at least two electrodes, wherein at least one electrode is positioned in the microfluidic trapping region and wherein the electrical potential provides a force attracting the reaction product molecules toward the microfluidic trapping region;
- concentrating the reaction product molecules in the microfluidic trapping region in front of, at a surface of and/or within the semi-permeable membrane;
- and
- detecting an amount of the concentrated reaction product molecules in the microfluidic trapping region in front of, at the surface of and/or within the semi-permeable membrane.
5 more elements
A method for detecting a target analyte, the method comprising the steps of:
- providing a microfluidic device comprising a binding surface in fluid communication with a microfluidic trapping region and at least two electrodes, wherein the microfluidic trapping region comprises a semi-permeable membrane, wherein the microfluidic trapping region comprises a semi-permeable membrane and wherein the binding surface comprises binding molecules capable of selectively binding to the target analyte;
- providing to the binding surface a first solution comprising target analyte molecules, wherein at least a number of the target analyte molecules selectively bind to the binding molecules of the binding surface, thereby creating an analyte activated binding surface;
- providing to the analyte activated binding surface a second solution comprising catalyst molecules, wherein at least a number of the catalyst molecules further binds directly or indirectly to the analyte activated binding surface, thereby creating a catalytic binding surface;
- providing to the catalytic binding surface a third solution comprising substrate molecules, the substrate molecules comprising a chromophore, wherein at least a number of the substrate molecules undergo a catalytic chemical reaction at the catalytic binding surface, thereby producing reaction product molecules, the reaction product molecules comprising the chromophore in the same or a substantially unaltered form as in the substrate molecules and wherein an ionic charge of the reaction product molecules is different from an ionic charge of the substrate molecules;
- transporting at least a number of the reaction product molecules into the trapping region by applying an electrical potential between the at least two electrodes, wherein at least one electrode is positioned in the microfluidic trapping region and wherein the electrical potential provides a force attracting the reaction product molecules toward the microfluidic trapping region;
- concentrating reaction product molecules in the microfluidic trapping region in front of, at a surface of and/or within the semi-permeable membrane;
- and
- detecting an amount of concentrated reaction product molecules in the microfluidic trapping region in front of, at a surface of and/or within the semi-permeable membrane.
5 more elements
Dependencies are inferred from the claim text.