Hybrid nanopore sensors

Abstract

The disclosure provides detection apparatus having one or more nanopores, methods for making apparatus having one or more nanopore and methods for using apparatus having one or more nanopores. Uses include, but are not limited to detection and sequencing of nucleic acids.

1 .- 104 . (canceled) 
     
     
         105 . A method of making a detection apparatus, comprising:
 (a) providing a solid support comprising an array of solid state nanopores;   (b) providing a plurality of lipid nanodiscs, wherein each lipid nanodisc comprises a protein nanopore inserted in a lipid bilayer; and   (c) contacting the plurality of lipid nanodiscs with the array of solid state nanopores so that each solid state nanopore becomes covered by one lipid nanodisc.   
     
     
         106 . The method of  claim 105 , wherein (b) comprises incubating the protein nanopores with phospholipids in the presence of a detergent; and removing the detergent from the phospholipids to form the lipid nanodiscs. 
     
     
         107 . The method of  claim 105 , wherein (c) comprises contacting the solid state nanopores with an amount of the plurality of lipid nanodiscs that exceeds an amount of the solid state nanopores. 
     
     
         108 . The method of  claim 105 , wherein each lipid nanodisc in the plurality of lipid nanodiscs is stabilized with a membrane scaffold protein (MSP). 
     
     
         109 . The method of  claim 105 , wherein each lipid nanodisc in the plurality of lipid nanodiscs comprises no more than one protein nanopore. 
     
     
         110 . The method of  claim 105 , wherein each lipid nanodisc in the plurality of lipid nanodiscs comprises charged tethers attached to the lipid bilayer or to the protein nanopore. 
     
     
         111 . The method of  claim 110 , wherein the charged tethers comprise nucleic acids. 
     
     
         112 . The method of  claim 110 , wherein (c) comprises electrically attracting the plurality of lipid nanodiscs to the solid state nanopores. 
     
     
         113 . The method of  claim 105 , further comprising (d) coupling the plurality of lipid nanodiscs with the array of solid state nanopores. 
     
     
         114 . The method of  claim 113 , wherein the coupling is via a silane, a cholesterol, or a cholesterol derivative. 
     
     
         115 . The method of  claim 105 , wherein the plurality of lipid nanodiscs are non-contiguous from each other on a surface of the solid support. 
     
     
         116 . The method of  claim 105 , wherein the plurality of lipid nanodiscs form seals for at least 50% of the solid state nanopores. 
     
     
         117 . The method of  claim 105 , wherein the apparatus comprises a cis reservoir in contact with the array of solid state nanopores and a trans reservoir in contact with the array of solid state nanopores, and wherein the cis reservoir and the trans reservoir comprise electrodes to apply a current through apertures formed by the protein nanopores. 
     
     
         118 . The method of  claim 105 , wherein the apparatus comprises an amplifier configured to amplify electrical signals generated at the protein nanopores. 
     
     
         119 . A method of making a nanopore detection apparatus, comprising:
 (a) providing a plurality of lipid nanodiscs, wherein each lipid nanodisc comprises a protein nanopore inserted in a lipid bilayer; and   (b) coupling each lipid nanodisc to a solid state nanopore by contacting the plurality of lipid nanodiscs to a solid support comprising an array of solid state nanopores.   
     
     
         120 . The method of  claim 119 , wherein (a) comprises incubating a plurality of protein nanopores with phospholipids in the presence of a detergent; and removing the detergent from the phospholipids to form the plurality of lipid nanodiscs. 
     
     
         121 . The method of  claim 119 , wherein (b) comprises contacting a plurality of solid state nanopores with an amount of the plurality of lipid nanodiscs that exceeds an amount of a plurality of solid state nanopores. 
     
     
         122 . The method of  claim 119 , wherein (b) comprises electrically attracting the plurality of lipid nanodiscs to a plurality of solid state nanopores.