Chemiresistive Biosensor for the Quantitative Detection of Human Cardiac Biomarker and a Process Thereof

• US 20160202250A1
• Filed: 01/08/2016
• Published: 07/14/2016
• Est. Priority Date: 01/09/2015
• Status: Abandoned Application

• Current Assignee: Council of Scientific & Industrial Research
• Original Assignee: Council of Scientific & Industrial Research
• Inventors: SHARMA, Vikash, TANWAR, Vinod Kumar, BIRADAR, Ashok Manikrao, Rajesh
• Application Number:
  US14/991,527
• Publication Number:
  US 20160202250A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 40/00 Manufacture or treatment of nanostructures
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C25D 13/02 with inorganic material
• CPC Class Codes: C25D 13/18 using modulated, pulsed, or reversing current
• CPC Class Codes: G01N 2333/4712 Muscle proteins, e.g. myosin, actin, protein
• CPC Class Codes: G01N 2333/805 Haemoglobins; Myoglobins
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/587 Nanoparticles

Abstract

The present invention disclosed a metal nanoparticles/single-walled carbon nanotube (MNP/SWCNT) hybrid based chemiresistive biosensor for the quantitative detection of human cardiac biomarkers troponin I (cTnI) and myoglobin (Mb). The highly specific cardiac-antibody, anti-cTnI (Ab-cTnI) or anti-Mb (Ab-Mb), was covalently immobilized to site-specific carboxyl groups on MNP anchored over SWCNT device. The biosensor device was characterized by the source-drain current-voltage measurements. The device performance was investigated with a change in conductance in SWCNT channel upon exposure to cTnI in human serum. MNP provided large surface area for high protein loading and improved electrical signal by inducing charge density in SWCNT, resulting in low level detection of cTnI and Mb with high sensitivity.

Claims

1. A label free chemiresistive biosensor device for quantitative detection of human cardiac biomarker, wherein cardiac biomarkers are selected from human cardiac troponin I (cTnI) and myoglobin(Mb), said biosensor device comprising aligned carbon nanotubes channel, bridging an interspaced gap between microfabricated gold source and drain electrode over silicon dioxide coated silicon wafer having gold electrode surface passivated with thiol molecules and the carbon nanotubes channel being functionalized with capped platinum nanoparticles through a bilinker 1-pyrenemethylamine hydrochloride and said platinum nanoparticles being covalently immobilized with cardiac biomarker specific protein antibody anti-cTnI (Ab-cTnI) or anti-Mb (Ab-Mb) and blocking non-specific binding sites with a blocking reagent.

2. The device as claimed in claim 1, wherein the carbon nanotube is single-walled carbon nanotubes (SWCNTs) having 80% p-type semiconducting characteristics.

3. The device as claimed in claim 1, wherein the interspaced gap between the microfabricated gold source and drain electrodes is in the range of 1-5 μ

m.

4. The device as claimed in claim 1, wherein the platinum nanoparticles is capped with molecules having functional groups selected from the group consisting of carboxyl, amino and hydroxyl.

5. The device as claimed in claim 4, wherein the platinum nanoparticles is capped with mercaptopropionic acid.

6. The device as claimed in claim 1, wherein the size of the platinum nanoparticles is in the range of 2-20 nm.

7. The device as claimed in claim 6, wherein the size of the platinum nanoparticles in the range of 2-10 nm.

8. The device as claimed in claim 1, wherein the cardiac biomarker used is selected from cTnI and Mb. wherein the biosensor device is selective and sensitive to human cardiac cTnI in the range of 0.001 ng to 10.0 ng mL⁻

1 in normal human serum.

9. The device as claimed in claim 1, wherein the biosensor device is selective and sensitive to cTnI with a sensitivity of about 11% to 75% change in resistance over the concentration range of 0.001 ng to 10.0 ng mL⁻

1 cTnI in human serum.

10. The device as claimed in claim 1 wherein the biosensor device is sensitive to myoglobin in the range of 0.03 ng to 1000 ng mL⁻

1 in phosphate buffer saline. wherein the lowest detection limit is 0.03 ng mL<sup>−

1</sup> for myoglobin.

11. The device as claimed in claim 1, wherein the thiol molecules are selected from the group consisting of 6-mercapto-1-haxanol and ethane hexanol.

12. The device as claimed in claim 1, wherein the blocking reagent is selected from the group consisting of bovine serum albumin, ethylamine and Polyoxyethylene (20) sorbitan monolaurate.

13. A process for the preparation of a label free chemiresistive biosensor device comprising the steps of:

i) bridging interspaced gap in between microfabricated gold electrode prepared over the silicon dioxide coated silicon substrate by electrophoretically aligning the single walled-carbon nanotubes, followed by annealing at a temperature of 200-400°

C. under reduced environment of a gaseous mixture of 5-10% hydrogen in nitrogen gas, washing with distilled water and drying under N₂ gas flow to obtain a aligned single walled carbon nanotubes device (SWCNTs),

ii) treating the aligned single walled carbon nanotubes, as obtained in step (i) with a bilinker 1-pyrenemethylamine hydrochloride having terminal amino group in solvent for a period of 1-3 h and passivating gold source and drain electrodes by treating the carbon nanotubes with thiol molecule in dimethyl formamide for a period of 1-5 h, followed by washing successively with dimethyl formamide and distilled water and dried under N₂ gas flow to obtain bilinker modified single walled-carbon nanotubes,

iii) functionalizing bilinker modified single walled-carbon nanotubes, as obtained in step (ii) with carboxyl capped platinum nanoparticles (PtNP) by exposing the modified SWCNT-device to an aqueous solution of metal nanoparticles containing N-(3-dimethylaminopropyl)-N-ethyl carbodiimide hydrochloride and N-hydroxy succinimide for 1-5 h, rinsing thoroughly with double distilled water and drying under N₂ gas flow to obtain the PtNPs/SWCNT hybrid device,

iv) immobilizing cardiac biomarker specific protein antibody on PtNPs/SWCNT hybrid device, as obtained in step (iii) by exposing the hybrid device to antibody solution in phosphate buffer saline, at pH 7.0-7.6, at a temperature of 4-10°

C., for a period of 10-24 h, followed by washing with double distilled water and drying under N₂ gas flow and passivating the device with a blocking reagent to obtain the chemiresistive biosensor device.

14. The process as claimed in claim 13, wherein thiol molecule is selected from the group consisting of 6-mercapto-1-haxanol and ethane hexanol.

15. The process as claimed in claim 14, wherein solvent is selected from the group consisting of dimethyl formamide and water or mixture thereof

16. The process as claimed in claim 14, wherein blocking reagent is selected from the group consisting of bovine serum albumin, ethylamine and Polyoxyethylene (20) sorbitan monolaurate.