Apparatus and methods for detecting molecules and binding energy

Abstract

The present disclosure provides apparatuses and methods for analyzing the presence of a target analyte. The apparatuses and methods of the present disclosure can be operated in a multiplexed format to perform various assays of clinical significance.

1 - 51 . (canceled) 
     
     
         52 . A method, comprising:
 providing (i) a sensor surface comprising a thiol group, (ii) a target molecule comprising an azide group, and (iii) a linker molecule comprising a maleimide moiety and a strained triple bond; and   reacting the linker molecule with the sensor surface and the target molecule such that (a) the azide group of the target molecule and the strained triple bond of the linker molecule undergo cycloaddition, and (b) the thiol group of the sensor surface forms a covalent bond with the maleimide moiety of the linker molecule, thereby coupling the target molecule to the sensor surface via the linker molecule.   
     
     
         53 . The method of  claim 52 , wherein the strained triple bond is part of a dibenzocyclooctyne (DIBO) group. 
     
     
         54 . The method of  claim 52 , wherein the reacting the linker molecule with the sensor surface and the target molecule is not catalyzed by copper. 
     
     
         55 . The method of  claim 52 , wherein the linker molecule is reacted simultaneously with the sensor surface and the target molecule. 
     
     
         56 . The method of  claim 52 , wherein the linker molecule is reacted with the sensor surface prior to being reacted with the target molecule. 
     
     
         57 . The method of  claim 52 , wherein the linker molecule is reacted with the sensor surface subsequent to being reacted with the target molecule. 
     
     
         58 . The method of  claim 52 , wherein the target molecule is a nucleic acid molecule. 
     
     
         59 . The method of  claim 52 , wherein the target molecule is an antibody. 
     
     
         60 . The method of  claim 59 , wherein the azide group of the antibody is not in proximity to a binding region of the antibody. 
     
     
         61 . The method of  claim 59 , wherein the azide group of the antibody is located in a constant region of the antibody. 
     
     
         62 . The method of  claim 59 , wherein the azide group of the antibody is not located in a variable region of the antibody. 
     
     
         63 . The method of  claim 60 , wherein the azide group is added to the antibody at least in part by removing terminal galactose residues of terminal N-acetylglucosamine (GlcNAc) residues using the enzyme beta-galactosidase and incorporating a N-azidoacetylgalactosamine-tetraacylated (GalNAz) which contains the azide group into the antibody using a beta-galactosyltransferase (GalT), thereby adding the azide group to antibody. 
     
     
         64 . The method of  claim 61 , wherein the azide group is added to the antibody at least in part by removing terminal galactose residues of terminal N-acetylglucosamine (GlcNAc) residues using the enzyme beta-galactosidase and incorporating a N-azidoacetylgalactosamine-tetraacylated (GalNAz) which contains the azide group into the antibody using a beta-galactosyltransferase (GalT), thereby adding the azide group to antibody. 
     
     
         65 . The method of  claim 62 , wherein the azide group is added to the antibody at least in part by removing terminal galactose residues of terminal N-acetylglucosamine (GlcNAc) residues using the enzyme beta-galactosidase and incorporating a N-azidoacetylgalactosamine-tetraacylated (GalNAz) which contains the azide group into the antibody using a beta-galactosyltransferase (GalT), thereby adding the azide group to antibody. 
     
     
         66 . The method of  claim 63 , wherein the sensor surface is part of a sensor that comprises a plurality of electrodes capable of applying an electric field. 
     
     
         67 . The method of  claim 64 , wherein the sensor surface is part of a sensor that comprises a plurality of electrodes capable of applying an electric field. 
     
     
         68 . The method of  claim 65 , wherein the sensor surface is part of a sensor that comprises a plurality of electrodes capable of applying an electric field. 
     
     
         69 . The method of  claim 52 , wherein the reacting the linker molecule is at an efficiency of at least about 20%.