Apparatuses and methods for detecting molecules and binding energy

Abstract

The present disclosure provides apparatuses and methods for analyzing the presence of charged analytes and/or the binding force between charged analytes and a capture probe. The apparatuses and methods of the present disclosure can be operated in a multiplexed format to perform various assays of clinical significance for example.

1 .- 57 . (canceled) 
     
     
         58 . An apparatus for detecting a presence of an analyte, comprising:
 a fluidic chamber adapted to contain an electrolyte;   two or more electrodes capable of generating an electric field within the fluidic chamber;   at least one surface comprising an immobilized capture probe capable of binding the analyte, wherein the at least one surface is between the two or more electrodes and in contact with the fluidic chamber, and wherein the at least one surface comprises an electrically conducting layer and an electrically insulating layer; and   at least one field confining feature proximal to the at least one surface, which at least one field confining feature is capable of concentrating an electric field surrounding the at least one field confining feature to a strength of at least about 10 3  volts per meter (V/m).   
     
     
         59 . The apparatus of  claim 58 , wherein the analyte is a charged analyte. 
     
     
         60 . The apparatus of  claim 58 , wherein the at least one field confining feature is capable of concentrating the electric field surrounding the at least one field confining feature to a strength of less than about 10 5  V/m. 
     
     
         61 . The apparatus of  claim 58 , further comprising a controller operably coupled to the two or more electrodes, wherein the controller is programmed to apply a voltage across the two or more electrodes that is sufficient to generate an applied force on the analyte. 
     
     
         62 . (canceled) 
     
     
         63 . The apparatus of  claim 58 , wherein the voltage is sufficient to generate an applied force of at least about 20 pN on the analyte. 
     
     
         64 . The apparatus of  claim 58 , wherein the immobilized capture probe is capable of binding to the analyte to form a probe-analyte complex on the at least one surface. 
     
     
         65 . The apparatus of  claim 64 , wherein the immobilized capture probe is capable of binding to the analyte via non-covalent interaction(s). 
     
     
         66 . The apparatus of  claim 65 , wherein the non-covalent interaction(s) are disruptable through an applied force generated upon the application of a voltage across the two or more electrodes. 
     
     
         67 . The apparatus of  claim 64 , wherein the immobilized capture probe is proximal to the at least one field confining feature. 
     
     
         68 . (canceled) 
     
     
         69 . The apparatus of  claim 58 , wherein the immobilized capture probe is an antibody. 
     
     
         70 . (canceled) 
     
     
         71 . The apparatus of  claim 58 , wherein the immobilized capture probe is a nucleic acid molecule. 
     
     
         72 .- 74 . (canceled) 
     
     
         75 . The apparatus of  claim 58 , further comprising a controller operably coupled to the two or more electrodes, wherein the controller is programmed to apply a potential difference of less than about 100 volts (V) between the two or more electrodes. 
     
     
         76 . The apparatus of  claim 58 , wherein the at least one surface provides a wall of the fluidic chamber and the field confining features is an orifice in the surface. 
     
     
         77 . The apparatus of  claim 58 , wherein the field confining feature comprises an orifice in the at least one surface. 
     
     
         78 .- 79 . (canceled) 
     
     
         80 . The apparatus of  claim 77 , wherein the orifice is an indentation, a well, a pore, a channel, a gap, or a slit. 
     
     
         81 .- 85 . (canceled) 
     
     
         86 . The apparatus of  claim 58 , wherein the field confining feature has a sharp edge. 
     
     
         87 . (canceled) 
     
     
         88 . The apparatus of  claim 58 , wherein the electrically conducting layer is in communication with the electrolyte. 
     
     
         89 .- 90 . (canceled) 
     
     
         91 . The apparatus of  claim 58 , further comprising a sensing electrode configured to measure a charge signal associated with the presence of the analyte. 
     
     
         92 . The apparatus of  claim 91 , wherein the sensing electrode is proximal to the at least one field confining feature. 
     
     
         93 .- 97 . (canceled) 
     
     
         98 . The apparatus of  claim 58 , wherein the at least one surface is part of an array of sensors, wherein each sensor of the array comprises a field confining feature and a plurality of immobilized capture probes. 
     
     
         99 .- 109 . (canceled)