Biological membrane-based sensor

Abstract

Provided herein is a biosensor suitable for use in measuring membrane fluidity or membrane permeability. The biosensor is formed of a solid substrate having a lipid bilayer compatible surface and a multi-lamellar lipid membrane structure localized on the lipid bilayer compatible surface. The multi-lamellar lipid membrane structure can be derived from a biological cell further comprising one or more synthetic lipids. An electrode forming all or part of the lipid bilayer compatible surface may be used to detect disruptions in the lipid membrane structure and hemolytic activity in a test sample.

1 . A biosensor comprising:
 a solid substrate having a lipid bilayer compatible surface; and   a multi-lamellar lipid membrane structure localized on the lipid bilayer compatible surface, the multi-lamellar lipid membrane structure being prepared from red blood cells or red blood cell ghosts and comprising one or more synthetic lipids.   
     
     
         2 . The biosensor of  claim 1 , wherein the one or more synthetic lipids are selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS), 1,2-dimyristoyl-sn-glycero-3-phospho-L-serine (DMPS), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG). 
     
     
         3 . The biosensor of  claim 1 , wherein the one or more synthetic lipids comprises a phospholipid, wherein the phospholipid is 18:1, 18:0, 14:0, or 16:0. 
     
     
         4 . The biosensor of  claim 1 , wherein the one or more synthetic lipids comprises a phosphatidylcholine (PC) and/or a phosphatidylethanolamine (PE). 
     
     
         5 . The biosensor of  claim 4 , wherein the PC is selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC). 
     
     
         6 . The biosensor of  claim 4 , wherein the concentration of the PC and/or the PE in the multi-lamellar lipid membrane structure is between about 10 mass % to about 90 mass %, between about 20 mass % to about 80 mass %, between about 30 mass % to about 70 mass %, or between about 40 mass % to about 60 mass %, or between about 45 mass % to about 55 mass %. 
     
     
         7 . The biosensor of  claim 6 , wherein the concentration of the PC and/or the PE in the multi-lamellar lipid membrane structure is about 10 mass %, about 20 mass %, about 50 mass %, about 80 mass % or about 90 mass %. 
     
     
         8 . The biosensor of  claim 1 , wherein the one or more synthetic lipids comprises a phosphatidylglycerol (PG) and/or a phosphatidylserine (PS). 
     
     
         9 . The biosensor of  claim 8 , wherein the PS is selected from the group consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) and 1,2-dimyristoyl-sn-glycero phospho-L-serine (DMPS). 
     
     
         10 . The biosensor of  claim 8 , wherein the PG comprises 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG). 
     
     
         11 . The biosensor of  claim 8 , wherein the concentration of the PG and/or the PS in the multi-lamellar lipid membrane structure is between about 10 mass % to about 50 mass %, between about 20 mass % to about 40 mass %, or between about 25 mass % to about 35 mass %. 
     
     
         12 . The biosensor of  claim 11 , wherein the concentration of the PG and/or the PS in the multi-lamellar lipid membrane structure is about 10 mass %, about 20 mass %, or about 50 mass %. 
     
     
         13 . The biosensor of  claim 1 , wherein the multi-lamellar lipid membrane structure further comprises one or more lipids conjugated with polyethylene glycol (PEGylated lipids). 
     
     
         14 . The biosensor of  claim 13 , wherein the PEG is PEG 350, PEG550, PEG 750, PEG1000, PEG2000, PEG3000, or PEG5000. 
     
     
         15 . The biosensor of  claim 13 , wherein the one or more PEGylated lipids comprises PEGylated phosphatidylethanolamine (PE). 
     
     
         16 . The biosensor of 13, wherein the one or more PEGylated lipids comprises a functionalized PEGylated lipid, the functionalized PEGylated lipid comprises a functionalized group selected from the group consisting of maleimide, biotin and amine. 
     
     
         17 . The biosensor of  claim 15 , wherein the PEGylated PE is 18:1, 18:0, 14:0, or 16:0. 
     
     
         18 . The biosensor of  claim 13 , wherein the one or more PEGylated lipids comprises PEGylated cholesterol. 
     
     
         19 . The biosensor of  claim 13 , wherein the polyethylene glycol is methoxy polyethylene glycol. 
     
     
         20 . The biosensor of  claim 13 , wherein the concentration of the one or more synthetic lipids is up to about 10 mass % and the concentration of the PEGylated lipids in the multi-lamellar lipid membrane structure is up to about 10 mass %. 
     
     
         21 . A biosensor comprising:
 a solid substrate having a lipid bilayer compatible surface; and   a multi-lamellar lipid membrane structure localized on the lipid bilayer compatible surface, the multi-lamellar lipid membrane structure comprises two or more synthetic lipids comprising a phosphatidylethanolamine (PE), a phosphatidylglycerol (PG), a phosphatidylcholine (PC), a phosphatidylserine (PS), a cholesterol, and/or a cardiolipin (CL).   
     
     
         22 . The biosensor of  claim 21 , wherein the PE comprises 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and/or 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE). 
     
     
         23 . The biosensor of  claim 21 , wherein the PG comprises 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG) and/or1,2-dimyristoyl-sn-glycero-3--phospho-(1′-rac-glycerol) (DMPG). 
     
     
         24 . The biosensor of  claim 21 , wherein the PC comprises 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). 
     
     
         25 . The biosensor of  claim 21 , wherein the PS comprises 1,2-dimyristoyl-sn-glycero-3-phospho-L-serine (DMPS). 
     
     
         26 . The biosensor of  claim 21 , wherein the two or more synthetic lipids are PE, PG and CL, wherein the concentration of the PE in the multi-lamellar lipid membrane structure is between about 67 mass % to about 71 mass %, the concentration of the PG is in the multi-lamellar lipid membrane structure is between about 23 mass % to about 27 mass %, and the concentration of the CL in the multi-lamellar lipid membrane structure is between about 3 mass % to about 10 mass %. 
     
     
         27 . The biosensor of  claim 21 , wherein the two or more synthetic lipids are PE and CL, wherein the concentration of the PE in the multi-lamellar lipid membrane structure is about 90 mass % and the concentration of the CL in the multi-lamellar lipid membrane structure is about 10 mass %. 
     
     
         28 . The biosensor of  claim 21 , wherein the two or more synthetic lipids are PE and PG, wherein the concentration of the PE in the multi-lamellar lipid membrane structure is about 78 mass % and the concentration of the PG in the multi-lamellar lipid membrane structure is about 22 mass %. 
     
     
         29 . The biosensor of  claim 21 , wherein the two or more synthetic lipids are PG and CL, wherein the concentration of the PG in the multi-lamellar lipid membrane structure is about 80 mass % and the concentration of the CL in the multi-lamellar lipid membrane structure is about 20 mass %. 
     
     
         30 . The biosensor of  claim 21 , wherein the two or more synthetic lipids are PC, PS, PE and cholesterol. 
     
     
         31 . The biosensor of  claim 30 , wherein the PC is 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), the PS is 1,2-dimyristoyl-sn-glycero-3-phospho-L-serine (DMPS), and the PE is 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE). 
     
     
         32 . The biosensor of  claim 30 , wherein the mass ratio of PC:PS:PE:cholesterol is 2:1:1:0.2 or 2:1:1:0. 
     
     
         33 . A method of detecting membrane disruption activity in a sample, the method comprising: 
       (a) contacting the sample with the biosensor of  claim 1 ; and 
       (b) detecting a change in the multi-lamellar lipid membrane structure in response to the sample. 
     
     
         34 . The method of  claim 33 , wherein the biosensor comprises at least one electrode and detecting the change in the multi-lamellar lipid membrane structure comprises voltammetry, cyclic voltammetry, chronoamperometry, differential multi pulse voltammetry, double potential pulse techniques or additive differential pulse voltammetry. 
     
     
         35 . A method of making the biosensor of  claim 1 , the method comprising:
 (a) preparing an aqueous solution of hybrid membranes comprising membranes prepared from red blood cells or red blood cell ghosts and the one or more synthetic lipids; and   (b) generating the multi-lamellar lipid membrane structure localized on the lipid bilayer compatible surface by successively providing the aqueous solution of hybrid membranes on the lipid bilayer compatible surface.   
     
     
         36 . The method of  claim 35 , wherein the aqueous solution of hybrid membranes is successively provided on the lipid bilayer compatible surface from 2 to 8 times, preferably 5 times. 
     
     
         37 . The method of  claim 35 , wherein after each providing of the aqueous solution of hybrid membranes to the lipid bilayer compatible surface, the method further comprises allowing the hybrid membranes to attach to the biosensor, and performing annealing to improve stability and reduce defects in the multi-lamellar lipid membrane structure.