TISSUE IMPLANTABLE MICROBIOSENSOR

• US 20130324820A1
• Filed: 12/01/2011
• Published: 12/05/2013
• Est. Priority Date: 12/01/2010
• Status: Active Grant

• Current Assignee: Pinnacle Technology, Inc.
• Original Assignee: Pinnacle Technology, Inc.
• Inventors: Petillo, Peter A., Wilson, George S., Wu, Judy Z., Aillon, Daniel V., Richter, Mark L., Johnson, David A.
• Granted Patent: US 9,468,402 B2
• Time in Patent Office: Days
• US Class Current: 600/345
• CPC Class Codes: A61B 2562/12 Manufacturing methods specially adapted for producing sensors for in-vivo measurements
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/14865 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
• CPC Class Codes: C25D 7/00 Electroplating characterised by the article coated

Abstract

A biosensor system comprises a capillary substrate, conductive electrode, and a plurality of nanoparticles having an enzyme deposited thereon formed in a cavity at one end of the capillary substrate. The substrate may have an optional reinforcing layer (which may be conductive or non-conductive) and optional insulating layer thereon. A cannula having an optional conductive layer, insulating layer, and reference electrode may also form part of the system.

Claims

1. A tissue implantable biosensor system comprising:

a capillary substrate;

a conductive electrode within said capillary substrate, said capillary substrate having a cavity at one end;

a plurality of nanoparticles having an enzyme immobilized thereon deposited in said cavity; and

a reference electrode.

2. The tissue implantable biosensor system of claim 1, wherein said capillary substrate has an exterior surface coated with a reinforcing material coating.

3. The tissue implantable biosensor system of claim 2, wherein said reinforcing material coating selected from the group consisting of a para-aramid polymer coating, carbon nanotube coating, and graphene coating.

4. The tissue implantable biosensor system of claim 3, wherein said reinforcing material forms an electric shield around said capillary substrate which reduces electrical noise and dielectric absorption effects in said system.

5. The tissue implantable biosensor system of claim 2, wherein said reinforcing material coating has a thickness from about 2 nm to about 100 nm.

6. The tissue implantable biosensor system of claim 2, wherein said reinforcing material coating has an overlying insulating layer.

7. The tissue implantable biosensor system of claim 1, wherein said conductive electrode comprises a metal or metal alloy wire which is sealed in the capillary substrate and etched to a depth to form said cavity.

8. The tissue implantable biosensor system of claim 7, wherein a resin seals said wire in said capillary substrate.

9. The tissue implantable biosensor system of claim 1, wherein said capillary substrate is housed in a cannula.

10. The tissue implantable biosensor system of claim 9, wherein said reference electrode is integrated in said cannula.

11. The tissue implantable biosensor system of claim 10, wherein said cannula is tapered.

12. The tissue implantable biosensor system of claim 10, wherein a conducting layer is formed on said cannula, said conducting layer forming an electric shield around said biosensor which reduces electrical noise and dielectric absorption effects in said system.

13. The tissue implantable biosensor system of claim 12, wherein an insulating layer overlies said conducting layer.

14. The tissue implantable biosensor system of claim 1, wherein said nanoparticlescomprise a nanoparticle core and a coating on said nanoparticle core, said coating comprising an activated group for covalent attachment of said enzyme to said nanoparticle core.

15. The tissue implantable biosensor system of claim 14, wherein said nanoparticle core comprises a metal, a metal alloy, or a combination thereof.

16. The tissue implantable biosensor system of claim 14, wherein said nanoparticle core is magnetic.

17. The tissue implantable biosensor system of claim 16, wherein said nanoparticle core is selected from the group consisting of iron, nickel, cobalt, yttrium, neodymium, samarium, and combinations thereof.

18. The tissue implantable biosensor system of claim 14, wherein said nanoparticle core is non-magnetic.

19. The tissue implantable biosensor system of claim 18, wherein said nanoparticle core is selected from group consisting of gold, silver, platinum, manganese, palladium, copper, zinc, ruthenium, rhodium, cadmium, stainless steel, iridium, and combinations thereof.

20. The tissue implantable biosensor system of claim 1, wherein said nanoparticles are from about 30 nm to about 200 nm in diameter.

21. The tissue implantable biosensor system of claim 14, wherein said coating is non-conducting.

22. The tissue implantable biosensor system of claim 14, wherein said coating comprises a carbohydrate polymer.

23. The tissue implantable biosensor system of claim 14, wherein said coating comprises a linear carbohydrate polymer.

24. The tissue implantable biosensor system of claim 14, wherein said coating comprises a polymer selected from the group consisting of glycosaminoglycans (including hyaluronan, chondroitin, chondroitin sulfate, heparin, heparan sulfate, dermatan, dermatan sulfate, keratan, keratan sulfate), mucopolysaccharides, celluloses, pectins, glucomannans, galactomannans, agars, alginic acids, chitins, inulins, glucans, amyloses, amylopectins, xanthan gums, dextrans, glycogens, arabinoxylans, and combinations thereof.

25. The tissue implantable biosensor system of claim 14, wherein said nanoparticle core is completely coated with said coating.

26. The tissue implantable biosensor system of claim 14, wherein said coating is from about 5 nm to about 10 nm thick.

27. The tissue implantable biosensor system of claim 14, wherein said enzyme is attached to said coating via a group selected from the group consisting of primary amines, carboxylates, thiols, aldehyde moieties, and non-covalently attachments using bioaffinities.

28. The tissue implantable biosensor system of claim 14, wherein said enzyme is an oxidase enzyme selected from the group consisting of malate oxidase, hexose oxidase, aryl-alcohol oxidase, L-gulonolactone oxidase, pyranose oxidase, L-sorbose oxidase, pyridoxine 4-oxidase (S)-2-hydroxy-acid oxidase, ecdysone oxidase, secondary-alcohol oxidase, 4-hydroxymandelate oxidase, long-chain-alcohol oxidase, thiamine oxidase, hydroxyphytanate oxidase, N-acylhexosamine oxidase, polyvinyl-alcohol oxidase, D-arabinono-1,4-lactone oxidase, vanillyl-alcohol oxidase, D-mannitol oxidase, alditol oxidase, choline dehydrogenase, gluconate 2-dehydrogenase, glucooligosaccharide oxidase, alcohol dehydrogenase, cellobiose dehydrogenase, aldehyde oxidase, glyoxylate oxidase, indole-3-acetaldehyde oxidase, aryl-aldehyde oxidase, retinal oxidase, abscisic-aldehyde oxidase, aldehyde ferredoxin oxidoreductase, indolepyruvate ferredoxin oxidoreductase, aldehyde dehydrogenase, dihydroorotate oxidase, dihydrouracil oxidase, tetrahydroberberine oxidase, tryptophan alpha,beta-oxidase, L-galactonolactone oxidase, acyl-CoA dehydrogenase, isoquinoline 1-oxidoreductase, quinaldate 4-oxidoreductase, D-aspartate oxidase, L-amino-acid oxidase, monoamine oxidase, pyridoxal 5′

-phosphate synthase, D-glutamate oxidase, ethanolamine oxidase, putrescine oxidase, cyclohexylamine oxidase, protein-lysine 6-oxidase, D-glutamate(D-aspartate) oxidase, L-lysine 6-oxidase, primary-amine oxidase, 7-chloro-L-tryptophan oxidase, N-methyl-L-amino-acid oxidase, non-specific polyamine oxidase, N8-acetylspermidine oxidase (propane-1,3-diamine-forming), N6-methyl-lysine oxidase, polyamine oxidase (propane-1,3-diamine-forming), N1-acetylpolyamine oxidase, spermine oxidase, L-pipecolate oxidase, dimethylglycine oxidase, polyamine oxidase, E-dihydrobenzophenanthridine oxidase, NAD(P)H oxidase, urate oxidase, aci-nitropropanoate oxidase, sulfite oxidase, methanethiol oxidase, prenylcysteine oxidase, L-ascorbate oxidase, 3-hydroxyanthranilate oxidase, rifamycin-B oxidase, superoxide dismutase, reticuline oxidase, lactate oxidase, D-amino acid oxidase, (S)-6-hydroxynicotine oxidase, (R)-6-hydroxynicotine oxidase, alcohol oxidase, pyruvate oxidase, glucose oxidase, glutamate oxidase, acyl coenzyme A oxidase, choline oxidase, glutathione sulfhydryl oxidase, glycerolphosphate oxidase, sarcosine oxidase, xanthine oxidase, oxalate oxidase, cholesterol oxidase, gamma-glutamyl-putrescine oxidase, GABA oxidase, histamine oxidase, diamine oxidase, nucleoside oxidase, L-lysine oxidase, L-aspartate oxidase, glycine oxidase, and galactose oxidase.

29. The tissue implantable biosensor system of claim 14, wherein said enzyme is an oxidase enzyme selected from the group consisting of lactate oxidase, D-amino acid oxidase, (S)-6-hydroxynicotine oxidase, (R)-6-hydroxynicotine oxidase, alcohol oxidase, pyruvate oxidase, glucose oxidase, glutamate oxidase, acyl coenzyme A oxidase, choline oxidase, glutathione sullhydryl oxidase, glycerolphosphate oxidase, sarcosine oxidase, xanthine oxidase, oxalate oxidase, cholesterol oxidase, gamma-glutamyl-putrescence oxidase, GABA oxidase, histamine oxidase, diamine oxidase, nucleoside oxidase, L-lysine oxidase, L-aspartate oxidase, glycine oxidase, and galactose oxidase.

30. The tissue implantable biosensor system of claim 1, wherein the number of capillary substrates is from about one to about eight.

31. A method for making a biosensor comprising:

providing at least one capillary substrate having a conductive electrode therein and having a cavity at one end; and

electrodepositing in said cavity a plurality of nanoparticles having an enzyme immobilized thereon.

32. The method of claim 31, wherein said nanoparticles comprise a nanoparticle core and a coating on said nanoparticle core, said coating having an activated group for covalent attachment of said enzyme to said nanoparticle.

33. A method of claim 32, wherein said nanoparticle core comprises a metal, a metal alloy, or a combination thereof.

34. A method of claim 32, wherein said nanoparticle core is magnetic.

35. The method of claim 34, wherein said nanoparticle core is selected from the group consisting of iron, nickel, cobalt, yttrium, neodymium, samarium, and combinations thereof.

36. A method of claim 32, wherein said nanoparticle core is non-magnetic.

37. The method of claim 36, wherein said nanoparticle core is selected from group consisting of gold, silver, platinum, manganese, palladium, copper, zinc, ruthenium, rhodium, cadmium, stainless steel, iridium, and combinations thereof.

38. The method of claim 31, wherein said nanoparticles are from about 30 nm to about 200 nm in diameter.

39. The method of claim 32, wherein said coating is non-conducting.

40. The method of claim 32, wherein said coating comprises a carbohydrate polymer.

41. The method of claim 32, wherein said coating comprises a linear carbohydrate polymer.

42. The method of claim 32, wherein said coating comprises a polymer selected from the group consisting of glycosaminoglycans (including hyaluronan, chondroitin, chondroitin sulfate, heparin, heparan sulfate, dermatan, dermatan sulfate, keratan, keratan sulfate), mucopolysaccharides, celluloses, pectins, glucomannans, galactomannans, agars, alginic acids, chitins, inulins, glucans, amyloses, amylopectins, xanthan gums, dextrans, glycogens, arabinoxylans, and combinations thereof.

43. The method of claim 32, wherein said nanoparticle core is completely coated with said coating.

44. The method of claim 32, wherein said coating is from about 5 nm to about 10 nm thick.

45. The method of claim 32, wherein said enzyme is covalently attached to said coating via a group selected from the group consisting of primary amines, carboxylates, thiols, aldehyde moieties, and non-covalent attachments using bioaffinities.

46. The method of claim 32, wherein said enzyme is an oxidase enzyme selected from the group consisting of malate oxidase, hexose oxidase, aryl-alcohol oxidase, L-gulonolactone oxidase, pyranose oxidase, L-sorbose oxidase, pyridoxine 4-oxidase (S)-2-hydroxy-acid oxidase, ecdysone oxidase, secondary-alcohol oxidase, 4-hydroxymandelate oxidase. long-chain-alcohol oxidase, thiamine oxidase, hydroxyphytanate oxidase, N-acylhexosamine oxidase, polyvinyl-alcohol oxidase, D-arabinono-1,4-lactone oxidase, vanillyl-alcohol oxidase, D-mannitol oxidase, alditol oxidase, choline dehydrogenase, gluconate 2-dehydrogenase, glucooligosaccharide oxidase, alcohol dehydrogenase, cellobiose dehydrogenase, aldehyde oxidase, glyoxylate oxidase, indole-3-acetaldehyde oxidase, aryl-aldehyde oxidase, retinal oxidase, abscisic-aldehyde oxidase, aldehyde ferredoxin oxidoreductase, indolepyruvate ferredoxin oxidoreductase, aldehyde dehydrogenase, dihydroorotate oxidase, dihydrouracil oxidase, tetrahydroberberine oxidase, tryptophan alpha,beta-oxidase, L-galactonolactone oxidase, acyl-CoA dehydrogenase, isoquinoline 1-oxidoreductase, quinaldate 4-oxidoreductase, D-aspartate oxidase, L-amino-acid oxidase, monoamine oxidase, pyridoxal 5′

-phosphate synthase, D-glutamate oxidase, ethanolamine oxidase, putrescine oxidase, cyclohexylamine oxidase, protein-lysine 6-oxidase, D-glutamate(D-aspartate) oxidase, L-lysine 6-oxidase, primary-amine oxidase, 7-chloro-L-tryptophan oxidase, N-methyl-L-amino-acid oxidase, non-specific polyamine oxidase, N8-acetylspermidine oxidase (propane-1,3-diamine-forming), N6-methyl-lysine oxidase, polyamine oxidase (propane-1,3-diamine-forming), N1-acetylpolyamine oxidase, spermine oxidase, L-pipecolate oxidase, dimethylglycine oxidase, polyamine oxidase, E-dihydrobenzophenanthridine oxidase, NAD(P)H oxidase, urate oxidase, aci-nitropropanoate oxidase, sulfite oxidase, methanethiol oxidase, prenylcysteine oxidase, L-ascorbate oxidase, 3-hydroxyanthranilate oxidase, rifamycin-B oxidase, superoxide dismutase, reticuline oxidase, lactate oxidase, D-amino acid oxidase, (S)-6-hydroxynicotine oxidase, (R)-6-hydroxynicotine oxidase, alcohol oxidase, pyruvate oxidase, glucose oxidase, glutamate oxidase, acyl coenzyme A oxidase, choline oxidase, glutathione sulfhydryl oxidase, glycerolphosphate oxidase, sarcosine oxidase, xanthine oxidase, oxalate oxidase, cholesterol oxidase, gamma-glutamyl-putrescine oxidase, GABA oxidase, histamine oxidase, diamine oxidase, nucleoside oxidase, L-lysine oxidase, L-aspartate oxidase, glycine oxidase, and galactose oxidase.

47. The method of claim 32, wherein said enzyme is an oxidase enzyme selected from the group consisting of lactate oxidase, D-amino acid oxidase, (S)-6-hydroxynicotine oxidase, (R)-6-hydroxynicotine oxidase, alcohol oxidase, pyruvate oxidase, glucose oxidase, glutamate oxidase, acyl coenzyme A oxidase, choline oxidase, glutathione sulfhydryl oxidase, glycerolphosphate oxidase, sarcosine oxidase, xanthine oxidase, oxalate oxidase, cholesterol oxidase, gamma-glutamyl-putrescence oxidase, GABA oxidase, histamine oxidase, diamine oxidase, nucleoside oxidase, L-lysine oxidase, L-aspartate oxidase, glycine oxidase, and galactose oxidase.

48. The method of claim 31, wherein said electrodepositing comprises applying a potential of from about 0.5 to about 1.0 V across said cavity.

49. The method of claim 31, wherein said potential is preferably applied for about 1 minute to about 30 minutes.

50. The method of claim 31, wherein said nanoparticles are dispersed in a solution having an ionic strength of from about 0.0001 M to about 0.001 M.

51. The method of claim 31, further comprising the step of electropolymerizing a monomer within said cavity to create a polymer intermingled with said nanoparticles.

52. The method of claim 51, wherein said monomer is selected from the group consisting of phenols and substituted phenols.

53. The method of claim 31, wherein said nanoparticles have a zeta potential such that said nanoparticles carry a majority of the current during said electrodepositing.

54. The method of claim 53, wherein the zeta potential ranges from about −

27 mV to about −

45 mV.

55. The method of claim 31, wherein said cavity has a volume ranging from about 0.001 pL to about 5 pL.

56. A tissue implantable cannula system comprising:

a cannula comprising a shaft configured to accommodate at least one biosensor;

an optional stylet to occupy the space within the shaft when the biosensor is absent;

a reference electrode integrated into said cannula;

an electrical circuit external to said cannula; and

a mechanism to connect said reference electrode to said external electrical circuit.

57. The tissue implantable cannula system of claim 56, wherein said shaft is about 1 μ

m to about 5 mm in diameter.

58. The tissue implantable cannula system of claim 56, wherein said shaft is about 0.1 mm to about 25 cm in length.

59. The tissue implantable cannula system of claim 56, wherein said shaft comprises a material selected from the group consisting of silver, gold, platinum, iridium, copper, iron, stainless steel, and combinations thereof.

60. The tissue implantable cannula system of claim 56, wherein said shaft comprises a layer of conducting and a layer of non-conducting material.

61. The tissue implantable cannula system of claim 56, wherein said shaft comprises at least two layers, wherein the inner-most layer comprises a non-conducting material and the outer-most layer comprises a conducting material, said conducting material acting as said integrated reference electrode.

62. The tissue implantable cannula system of claim 61, wherein said shaft comprises at least three layers.

63. The tissue implantable cannula system of claim 56, wherein said reference electrode is selected from the group consisting of silver, gold, platinum, iridium, copper, iron, stainless steel, and combinations thereof.

64. The tissue implantable cannula system of claim 56, wherein the shaft comprises at least one conducting layer that forms an electric shield around said biosensor to reduce electrical noise and dielectric absorption effects in the system.

65. The tissue implantable cannula system of claim 64, wherein said conducting layer acts as the integrated reference electrode.

66. The tissue implantable cannula system of claim 56, wherein:

said biosensor is selected from the group consisting of a capillary substrate, Pt wire, Pt/Ir wire, carbon fiber, metal wire, metal alloy wire and ceramic;

said biosensor may be sheathed in at least one layer of non-conducting material; and

said material is selected from the group consisting of glass, silica, Teflon, para-aramid polymer, carbon nanotubes, and graphene.