BML750:Point of care devices Technology

BIOSENSOR
To detect cardiavascular diseases in prior

-Rosy Painkra
Introduction
Traditional diagnosis methods for cardiovascular diseases are time
consuming and expensive since they are primarily conducted at
central clinical laboratories.

Faster and inexpensive diagnosis through pointofcare, labona

chip type systems is highly desirable.

Detection and quantification of biomarkers for acute myocardial

infraction (AMI) is critical in the diagnosis of cardiovascular
diseases.
Biorecognition element: A Biomarker
Biomarker-C protein(produced in liver and enters blood stream
produces inflammation in whole body)Concentration increases on
the onset of cardiac disease.

It can be combined with other biomarkers as Trponin,myoglobin etc

Measuring the levels of this protein can be helpful for predicting the
risks of cardiac diseases.

strong predictor of life threatening events such as AMI, giant cell

arteritis, arterial disease, stroke and cardiac arrest causing sudden
death .
Risk prediction based on level of CRP
Three different levels of CRP concentrations in human blood serum:
concentration less than 1 g/ml represents a low risk state

concentration between 1 to 3 g/ml is considered as average risk

concentration above 3 g/ml represents high risk

Electrochemical based biosensors
less expensive

faster, simpler

portable, accurate

flexible for multiplexing and

label free detection of a wide range of proteins, nucleic acids,

enzymes and other biomolecules.
Material for electrode:
carbon nanotubes (CNTs) (single and multi-walled), carbon nanofibers
(CNFs) graphene gold nano-wires, nano-structures increase:
sensitivity and detection limits for proteins, nucleic acids etc.
high conductivity, biocompatibility and ease of surface modification

Among these, vertically aligned CNFs(VACNFs), where each fiber is an

individual freestanding nanostructure acts as a nanoelectrode, have
been used successfully to construct biosensors.
Fabrication of Biosensor
A 4-inch silicon (100) wafer with 500 nm thermal oxide layer
consists of 30 devices.

Each device contains nine identical micro pads (electrodes)

chromium (Cr) layer was deposited by e-beam evaporation on

defined electrodes, contact pads and electrical interconnects

Then the electrodes (micropads) were spun coated with 400 nm

poly(methyl methacrylate) (PMMA) and patterned by using e-beam
current.
 A 10 nm Cr adhesion layer followed by 30 nm nickel catalyst layer
were further deposited on these e-beam patterned electrodes.

VACNFs were grown from these patterned Ni catalyst particles

attachment of the probe molecule anti-C-reactive protein to the

biosensor surface

This probe protein was used to capture C-reactive protein.

Growth of VACNF(vertically aligned carbon nano-fibre)

Method used:Plasma enhanced chemical vapor deposition

(PECVD)

Acetylene (C2H2) feedstock for carbon source diluted with ammonia.

Pressure of 6.3 mbar, Temperature:700 C and plasma power of 180 W.

A 15 minute deposition yields ~39000 free standing VACNFs spaced 1

m apart on each patterned electrode.
Electrochemical cell:

All electrochemical measurements were carried out using a

standard three electrodes electrochemical cell

The instrument was interfaced to a personal computer

The electrochemical cell consists of high quality platinum (Pt) wire

as counter electrode, saturated calomel electrode as reference
electrode and nine identical working electrodes of VACNF NEAs.
Quantitative analysis:
Cyclic voltammetry (CV)can beused for the CRP detection.

Insulation of the electrode's active surface area by presence of anti

CRP on the VACNF tips

Reduction of the amount of charged species (Fe [(CN)6]3-/4-)

diffusing towards the electrode causing reduction in the redox
currents.

Further decrease in redox current when CRP binds with anti-CRP

immobilized on the electrode proportional to the concentration of
the CRP target.
Electrochemical characterization of VACNF electrode recorded in electrolyte solution containing 5 mM
Fe[CN6]3-/4- in 1 M KCl.(CV)
Selected CV curves for bare and subsequent modified electrodes recorded in electrolyte solution containing 5
mM Fe[CN6]3-/4- in 1 M KCl;
A rapid and early diagnosis of CVD should involve for testing for a
definite marker with an early biomarker(CRP).

This biosensor can be linked with micro-fluidics for total analysis of the
CVD.
References:

1. Gupta, R. K. et al. Label-free detection of C-reactive protein using a carbon nanofiber

based biosensor. Biosens. Bioelectron. 59, 112119 (2014)
2. American Heart Association and the United States Center for Disease control