A Biosensor Device to Target Analytes in Situ, in Vivo, and/or in Real Time, and Methods of Making and Using the Same

• US 20160022185A1
• Filed: 03/11/2014
• Published: 01/28/2016
• Est. Priority Date: 03/11/2013
• Status: Active Grant

• Current Assignee: University of Toledo
• Original Assignee: University of Toledo
• Inventors: Goel, Vijay K., Agarwal, Anand K., Kim, Dong-Shik, Yoon, Do-Young, Lin, Boren, Feyzizarnagh, Hamid
• Granted Patent: US 10,849,540 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61B 2503/40 Animals
• CPC Class Codes: A61B 2562/125 characterised by the manufacture of electrodes
• CPC Class Codes: A61B 5/14546 for measuring analytes not otherwise provided for, e.g. ions, cytochromes
• CPC Class Codes: A61B 5/14735 comprising an immobilised reagent
• CPC Class Codes: A61B 5/4547 Evaluating teeth
• CPC Class Codes: G01N 27/3272 Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels optical biosensors G01N33/52
• CPC Class Codes: G01N 27/3276 being a hybridisation with immobilised receptors using a FET type sensor G01N27/4145; concerning the hybridisation C12Q1/68
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/56938 Staphylococcus
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

A biosensor device for the real-time detection of a target analyte includes a receptor component operatively connected to a transducer component which is adapted to interpret and transmit a detectable signal. The receptor component includes a sensing element capable of detecting and binding to at least one target analyte, and a self-assembled monolayer (SAM) layer. The SAM layer is positioned between and in contact with the sensing element and an electrode such that the sensing element, in the presence of the target analyte, causes a detectable signal capable of being transmitted to the electrode. The transducer component includes the electrode and microprocessor configured to screen noise and to pick up impedance change at a very low frequency range.

Claims

1. A biosensor for detecting the presence of a target analyte in a sample, the biosensor comprising:

a transducer component comprising an electrode operatively connected to a microprocessor, the microprocessor being adapted to receive, process and transmit a signal; and

,

a receptor component having;

i) a sensing element capable of detecting and binding to at least one target analyte present in a sample, and

ii) a self-assembled monolayer (SAM) layer, the SAM layer being positioned between and in contact with the sensing element and the electrode;

the transducer component and the receptor component being capable of being brought into direct contact with the sample in situ,

wherein the sensing element, in the presence of the target analyte present in the sample, causes a detectable signal capable of being transmitted to the electrode via the SAM layer.

2. The biosensor of claim 1, wherein the biosensor is capable of detecting the presence of the target analyte in real time.

3. The biosensor of claim 1, wherein the sensing element comprises at least one antibody capable of detecting at least one bacterial target analyte.

4. The biosensor of claim 1, wherein the sample comprises a fluid or tissue in a living organism.

5. The biosensor of claim 1, wherein the sample comprises a fluid or tissue in a living organism in vivo.

8. The biosensor of claim 1, wherein the sample comprises a food product.

9. The biosensor of claim 1, wherein the rate and degree of signal change correspond to the presence and concentration of the target analyte.

10. The biosensor of claim 1, wherein the presence of the target analyte is detected by impedance signal.

11. The biosensor of claim 1, wherein the detectable signal comprises a change in impedance as a function of frequency.

12. The biosensor device of claim 1, wherein the microprocessor is configured to screen noise and to pick up impedance change at a frequency range from about 1 Hz to about 10 Hz.

13. The biosensor of claim 1, wherein the presence of the target analyte is detected by amperometric signal.

14. The biosensor of claim 1, wherein the electrode comprises a micro-interdigitated gold electrode.

15. The biosensor of claim 1, wherein the detectable signal is displayed on the microprocessor through radio frequency identification (RFID).

16. The biosensor of claim 1, wherein the biosensor is integrated into a medical, dental, or veterinary device having a tissue-contacting surface.

19. The biosensor of claim 1, wherein the target analyte comprises a pathogen selected from the group consisting of:

Staphylococcus aureaus, methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus pyogenes, Streptococcus pneuomoniae, Streptococcus agalactiae, Staphylococcus epidermis, Staphylococcus saprophyticus, Escherichia coli, Legionella pneumophila, Pseudomonas aeruginosa, Enterococcus faecalis, Listeria, Cyclospora, Salmonella enteritidis, Helicobacter pylori, Tubercle bacillus (TB), other Bacillus, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium tetani, Sporohalobacter, Anaerobacter, Heliobacterium, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Cyanobacteria, green sulfur bacteria, Chloroflexi, purple bacteria, thermodesulfobacteria, hydrogenophilaceae, nitrospirae, Burkholderia cenocepacia, Mycobacterium avium, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium ulcerans, Lactobacillus, Lactococcus, Bordetella pertussis, Chlamydia pneumoniae, Chlamydia tracomatis, Chlamydia psittaci, Borrelia burgdorferi, Campylobacter jejuni, Francisella tularensis, Leptospira monocytogenes, Leptospira interrogans, Mycoplasma pneumoniae, Rickettsia rickettsii, Shigella sonnei, Traponema pallidum, Vibrio cholerae, Haemophilus influenzae, Neiserria meningitidis, Yersinia pestis, Bartonella henselae, Borrelia burgdorferi, Chlamydia psittaci, Dirofilaria immitis, Ehrlichia canis, Feline Calicivirus, Feline Coronavirus, Feline Herpesvirus, Feline Immunodeficiency Virus, Feline Leukemia Virus, Leptospira spp., Mycoplasma haemofelis, Panleukopenia Virus, Toxoplasma gondii, West Nile Virus, Canine Adenovirus, Canine Distemper Virus, Canine Herpesvirus, Bordetella bronchiseptica, Neospora Hughesi, Neospora Caninum, Anaplasma phagocytophilum, Rickettsia rickettsii, Anaplasma platys, Canine parainfluenza virus, Tritrichomonas foetus, Cryptosporidium spp., Cryptosporidium felis, Mycobacterium spp., Salmonella spp., Giardia spp, Taenia spp, Equine Herpes Virus, Equine Influenza A, Lawsonia intracellularis, Streptococcus equi, Equine Arteritis virus, Shigella spp., Yersinia enterocolitica, Rhodococcus equi, West Nile, Leptospira spp, Erysipelas rhusiopathiae, Bartonella, Coxiella, Pseudomonas sp., Pseudomonas pseudomallei, Pseudomonas mallei, Klebsiella, Salmonella sp., Enterococcus, Dolphin pox, seal pox, papilloma universal, papilloma manatee, canine adenovirus, influenza A and B, hepatitis A and B, Bovine enterovirus, Cosackivirus, encephalomyocarditis virus, Morbilliviruses, canine distemper virus, Bovine corona virus, Bovine rotavirus, universal herpes, echovirus, Aspergillus, Nocardia, Histoplasma, Blastomyces, Coccidioides immitis, Lacazia loboi, Saksenaea, and Aphophysomyces.

26. The biosensor of claim 1, wherein the receptor binds to a target analyte selected from the group consisting of:

pesticides, toxins, aflatoxins, arsenic, botulin, ciguatera toxin, cyanide, deoxynivalenol, dioxin, fungi, fumonisins, fusarium toxins, heavy metals, histadine, histamine, lead, marine toxins, mercury, mycotoxins, neurotoxin, nicotine, ochratoxin A toxins, patulin toxins, polychlorinated phenyls, pyrrolizidine alkaloids, ricin, scombrotoxins, shellfish toxin, tetrodotoxin, trichothecenes, zearelenone, and the combinations thereof.

27. The biosensor of claim 1, wherein the receptor binds to a target analyte selected from the group consisting of food allergens:

almond, egg, shellfish, gliadin, gluten, hazelnut, milk, peanut, soy residues, and combinations thereof.

28. The biosensor of claim 1, wherein the receptor binds to a target analyte selected from a virus, or portion thereof.

29. The biosensor of claim 1, wherein the receptor binds to a target analyte selected from a molecule, or portion thereof, that is a marker for a cancer.

30. The biosensor of claim 1, wherein the SAM comprises mercaptoproprionic acid (MPA), 11-mercaptoundecanoic acid (MUA), 1-tetradecanethiol (TDT), or dithiobios-N-succinimidyl propionate (DTSP).

38. A method of detecting a bacterial infection in a living organism, comprising:

placing the biosensor of claim 1 at least partially in or on the living organism sufficient to come into contact with any bacterial target analyte present in the living organism; and

,

detecting the presence of the bacterial target analyte when the biosensor device transmits the detectable signal.

39. The method of claim 38, wherein the biosensor device determines whether the bacterial target analyte is Gram-positive or Gram-negative, and the biosensor device transmits a signal indicating whether the bacterial target analyte is Gram-positive or Gram-negative.

40. The biosensor of claim 1, further comprising a membrane configured to protect the electrode, the membrane comprising an electrode mesh.

6-7. -7. (canceled)

17-18. -18. (canceled)

20-25. -25. (canceled)

31. (canceled)

32. A method of making a biosensor capable of detecting a target analyte in situ in a sample, comprising:

linking a sensing element comprising antibodies to an electrode via a self-assembled monolayer (SAM); and

operatively connecting a microprocessor to the electrode such that, when the sensing element binds to a target analyte present in situ in a sample, the microprocessor detects and transmits a signal.

33. The method of claim 32, further comprising the step of:

configuring the biosensor on a medical instrument from the group consisting of;

endoscopic imaging devices, harvesting devices, retractors, bone hooks, skin hooks, nerve hooks, tension devices, forceps, elevators, drill sleeves, osteotomes, spinal rongeurs, spreaders, gouges, bone files and rasps, bone awls, rib shears, trephines, suction tubes, taps, tamps, calipers, countersinks, suture passers, and probes.

36. The method of claim 32, further comprising using electrochemical impedance to detect and/or quantify the target analyte.

37. The method of claim 32, wherein the microprocessor includes an algorithm program capable of screening background noise and detecting an impedance signal that represents the presence and concentration of target analyte.

34-35. -35. (canceled)