Nanostructured separation and analysis devices for biological membranes

Abstract

The present invention provides a nonostructured device comprising a substrate including nanotroughs therein; and a lipid bilayer suspended on or supported in the substrate. A separation method is also provided comprising the steps of supporting or suspending a lipid bilayer on a substrate; wherein the subtrate comprises nanostructures and wherein the lipid bilayer comprises at least one membrane associated biomolecule; and applying a driving force to the lipid bilayer to separate the membrane associated biomolecule from the lipid bilayer and to drive the membrane associated biomolecule into the nanostructures.A fluidic device for separating particles according to size is provided including a fluidic channel, and a matrix comprising a plurality of protrusions within the fluidic channel, wherein the device provides a driving force to the particles being separated through the fluidic channel; and wherein a flow of the driving force from between the protrusions is divided unequally into a major flow component and a minor flow component, each component flowing between subsequent protrusions in the matrix, such that the average direction of the major flow component is not parallel to the average direction of the driving force, and, when particles are introduced into the matrix, particles having a size less than a predetermined critical size are transported generally in the average direction of the driving force, and particles having a size at least that of the critical size are transported generally in the average direction of the major flow component, thereby separating the particles according to size. Methods for separating particles including steps of separation based on size and affinity are also provided.

1. A nanostructured device comprising:
 a substrate including at least one nanotrough therein; and    a lipid bilayer suspended over said at least one nanotrough.    
     
     
       2. The nanostructured device of  claim 1 , wherein said lipid bilayer comprises a simple bilayer. 
     
     
       3. The nanostructured device of  claim 1 , wherein said lipid bilayer comprises a hybrid bilayer. 
     
     
       4. The nanostructured device of  claim 3 , wherein said hybrid bilayer comprises a self-assembled monolayer hybrid bilayer. 
     
     
       5. The nanostructured device of  claim 1 , wherein said at least one nanotrough is filled with at least one fluid. 
     
     
       6. The nanostructured device of  claim 1 , wherein said nanostructured device further comprises an array of nanostructures arranged so that said array has a gradient property. 
     
     
       7. The nanostructured device of  claim 1 , wherein said nanostructured device further comprises at least one nanostructured channel. 
     
     
       8. The nanostructured device of  claim 1 , wherein said substrate comprises Si. 
     
     
       9. The nanostructured device of  claim 1 , wherein said substrate comprises a semiconductor chip. 
     
     
       10. The nanostructured device of  claim 1 , wherein said nanostructured device comprises a biochip. 
     
     
       11. A nanostructured device comprising:
 a substrate including at least one nanotrough therein; and    at least one lipid bilayer supported in at least one of said at least one nanotroughs so as to allow biomolecules to pass from said at least one lipid bilayer into said at least one respective nanotrough.    
     
     
       12. The nanostructured device of  claim 11 , wherein said lipid bilayer comprises a simple bilayer. 
     
     
       13. The nanostructured device of  claim 11 , wherein said lipid bilayer comprises a hybrid bilayer. 
     
     
       14. The nanostructured device of  claim 13 , wherein said hybrid bilayer comprises a self-assembled monolayer hybrid bilayer. 
     
     
       15. The nanostructured device of  claim 11 , wherein said nanostructured device further comprises an array of nanostructures arranged so that said array has a gradient property. 
     
     
       16. The nanostructured device of  claim 11 , wherein said nanostructured device further comprises at least one nanostructured channel. 
     
     
       17. The nanostructured device of  claim 11 , wherein said substrate comprises Si. 
     
     
       18. The nanostructured device of  claim 11 , wherein said substrate comprises a semiconductor chip. 
     
     
       19. The nanostructured device of  claim 11 , wherein said nanostructured device comprises a biochip. 
     
     
       20. A separation method comprising the steps of:
 (a) supporting or suspending a lipid bilayer on a substrate; wherein said substrate comprises at least one nanostructure and wherein said lipid bilayer comprises at least one membrane associated biomolecule; and    (b) applying a driving force to said lipid bilayer to separate said at least one membrane associate biomolecule from said lipid bilayer and to drive said at least one membrane associated biomolecule within said lipid bilayer into said at least one nanostructure.    
     
     
       21. The method of  claim 20 , wherein said at least one nanostructure comprises at least one nanotrough. 
     
     
       22. The method of  claim 21 , wherein said at least one nanotrough is filled with at least one fluid. 
     
     
       23. The method of  claim 20 , wherein said at least one nanostructure comprises at least one channel. 
     
     
       24. The method of  claim 20 , wherein said at least one nanostructure further comprises at least one protrusion. 
     
     
       25. The method of  claim 20 , wherein said substrate comprises Si. 
     
     
       26. The method of  claim 20 , wherein said lipid bilayer comprises a simple bilayer. 
     
     
       27. The method of  claim 20 , wherein said lipid bilayer comprises a hybrid bilayer. 
     
     
       28. The method of  claim 27 , wherein said hybrid bilayer comprises a self-assembled monolayer hybrid bilayer. 
     
     
       29. The method of  claim 20 , wherein said at least one nanostructure comprises an array of nanostructures arranged so that said array has a gradient property. 
     
     
       30. The method of  claim 20 , wherein said at least one membrane associated biomolecule comprises a transmembrane protein. 
     
     
       31. The method of  claim 20 , wherein said at least one membrane associated biomolecule comprises a membrane phospholipid. 
     
     
       32. The method of  claim 20 , wherein said at least one membrane associated biomolecule comprises a lipophilic biomolecule. 
     
     
       33. The method of  claim 20 , wherein said driving force comprises electrophoresis. 
     
     
       34. The method of  claim 20 , wherein said driving force comprises an externally applied pressure. 
     
     
       35. The method of  claim 20 , wherein said driving force comprises capillarity. 
     
     
       36. The method of  claim 20 , wherein said driving force comprises diffusion. 
     
     
       37. The method of  claim 20 , wherein said driving force comprises osmosis. 
     
     
       38. A method for separating particles comprising:
 ( a )  providing a substrate and an array of structures arranged on said substrate, wherein said structures comprise protrusions fixed to said substrate in a predetermined pattern having a gradient property in a given direction across a plane of said substrate;      ( b )  a first separation step comprising separating said particles by size in two dimensions by flowing said particles through said array of structures in said given direction, the separating of said particles occurring at least in part by virtue of said pattern; and      ( c )  a second separation step comprising separating said particles by affinity.      
     
     
       39. The method of  claim 38 , wherein said separation based on affinity comprises affinity chromatography.  
     
     
       40. The method of  claim 39 , wherein said separation based on affinity chromatography comprises immunoaffinity.  
     
     
       41. The method of  claim 38 , wherein said gradient property is discrete.  
     
     
       42. The method of  claim 38 , wherein said structures create at least one channel.  
     
     
       43. The method of  claim 38 , wherein said substrate comprises Si.  
     
     
       44. The method of  claim 38 , wherein said substrate comprises silicon carbide.  
     
     
       45. The method of  claim 38 , wherein said particles are biomolecules.  
     
     
       46. The method of  claim 38 , wherein separating particles further comprises an additional separation step based on a mechanism selected from the group consisting of size exclusion, asymmetric diffusion, entropic trapping and hydrophobic interaction.  
     
     
       47. The method of  claim 38 , wherein said structures are surface modified.  
     
     
       48. The method of  claim 47 , wherein said surface modification comprises modification with poly- ethylene glycol.    
     
     
       49. The method of  claim 47 , wherein said surface modification comprises modification with one or more oligonucleotide probes.  
     
     
       50. The method of  claim 47 , wherein said surface modification comprises modification with a hydrophobic molecular or oligomeric species.  
     
     
       51. The method of  claim 47 , wherein said surface modification comprises modification with a chiral selector selected from the group consisting of cyclodextrins, macrocyclic antibiotics and crown ethers.  
     
     
       52. The method of  claim 38 , wherein separating particles further comprises applying a driving force to induce said flowing of particles.  
     
     
       53. The method of  claim 52 , wherein said driving force comprises externally applied pressure.  
     
     
       54. The method of  claim 52 , wherein said driving force comprises capillarity.  
     
     
       55. The method of  claim 52 , wherein said driving force comprises diffusion.  
     
     
       56. The method of  claim 52 , wherein said driving force comprises osmosis.