AN OVERVIEW OF RECENT DEVELOPMENTS IN NANOMATERIAL BASED ELECTROCHEMICAL BIOSENSORS

ARUN.S.SIDDARTH CENTRE FOR EDUCATION CENTRAL ELECTROCHEMICAL RESEARCH INSTITUTE (CSIR-CECRI) KARAIKUDI, TAMIL NADU

WHAT ARE BIOSENSORS?

Biosensor: biological sensing element connected to a

transducer to convert observed response into a measurable signal, whose magnitude is proportional to the concentration of a specific chemical or set of chemcials
Divided into several categories based on the

transduction process, such as electrochemical, optical, piezoelectric, and thermal/calorimetric biosensors

ELECROCHEMICAL BIOSENSORS
A molecular sensing device which couples a

biological recognition element to an electrode transducer. Amperometric or potentiometric transducers used. Potentiometric devices: biorecognition process converted into a potential signal Amperometric devices: monitors current associated with reduction or oxidation of electroactive species involved in the recognition process. Electrochemical biosensors- offer great promisefor biomedical applications.

NANOMATERIALS FOR ELECTROCHEMICAL BIOSENSORS

Metal nanoparticles: isolable particles between 1 and 50 nm in

size. Nanoparticles :physical, electronic and chemical properties different from those of bulk metals High surface-to-volume ratio and novel electron transport properties of nanostructures- strong influence of minor perturbations on electronic conductance. Extreme smallness of nanomaterials- allow packing a huge number of sensing elements onto small footprint of array device. Offer prospect of rapid and sensitive label-free bioelectronics detection, massive redundancy in nanosensor arrays 1D nanostructures like CNT, semiconductor or conducting polymer nanowires- particularly attractive for bioelectronic detection

ENZYME ELECTRODES.
Enzyme electrodes: used for monitoring wide range of

clinically or environmentally important substrates Establishment of satisfactory electrical communication between the active site of the enzyme and the electrode surface- major challenge [1] The redox center of most oxidoreductases is electrically insulated by protein shell. The enzyme cannot be oxidized or reduced at an electrode at any potential. The possibility of direct electron-transfer between enzymes and electrode surfaces could pave the way for superior reagentless biosensing devices

[1] Electroanalysis, 1997, 9, 661.

CNT BASED ENZYME ELECTRODES

Aligned CNT trees : prepared by self assembly Act as molecular wires to allow electrical communication

between the underlying electrode and redox proteins. linking of aligned reconstituted glucose oxidase (GOx) on the edge of SWCNT to an electrode surface demonstrated [2] Enzyme reconstitution on the end of CNT represents an extremely efficient approach for plugging an electrode into GOx. Interfacial electron transfer rate constant of 42 s-1 estimated for 50 nm long SWCNT. Edge plane defects at end caps- electron transfer at CNT

[2] Angew Chem. Int. Ed., 2004, 43, 2113.

Assembly of SWCNT electrically contacted glucose oxidase electrode: linking the reconstituted enzyme, on the edge of the FAD functionalized SWCNT, to the electrode surface.

OTHER NANOMATERIAL BASED ENZYME ELECTRODES

Catalytic properties of metal nanoparticles- facilitated

the electrical contact of redox centers of proteins with electrode surfaces. Gold nanoparticles: electron relays for alignment of glucose oxidase on conducting supports and wiring its redox center [3] Platinum nanoparticles deposited onto CNT: improvements in detection of the enzymatically-liberated peroxide species [4] Dispersed iridium nanoparticles (2 nm diameter) in graphite-like carbon: improved amperometric biosensing of glutamate [5]
[3] Science, 2003, 299, 1877 [4] Anal. Chem., 2010, 76, 1083. [5] Electroanalysis, 2007, 16, 54.

IN-VIVO GLUCOSE MONITORING

Pair of nanoelectrodes separated with a small (20-60

nm) gap connected by polyanaline/glucoseoxidase Film [6] IN-VIVO glucose monitoring

Electrodeposition within the channel between

electrodes- controllable route for preparing conducting polymer nanowire enzyme sensors [7]

[6] NanoLett, 2006, 4, 1785. [7] NanoLett, 2007, 4, 1237.

ELECTROCHEMICAL IMMUNOASSAYS
Abnormal concentrations of certain proteins can

indicate presence of cancers. Highly sensitive enzyme electrochemical Immunoassays developed [8] Rely on labeling of the antibody (or antigen) with an enzyme which acts on substrate and generate an electroactive product detected amperometrically. Possible to use metal markers and redox tags for electronic transduction of antigenantibody interactions in addition to enzyme labels.
[8] Trends Anal. Chem. 21, 213219.

NANOMATERIALS FOR IMMUNOASSAYS

One-dimensional (1D) nanostructures like

semiconductor or conducting-polymer nanowires (NW) extremely attractive for designing high-density protein arrays. High surface-to-volume ratio and novel electron transport properties and influence of minor perturbations on electronic conductance indicate great promise for label-free real-time protein detection Possible to pack large number of antibody-functionalized NW onto a remarkably small footprint of an array device [9] Great promise for assays of multiple disease markers in ultrasmall sample volumes.
[9] Science ,300, 1115.

NANOWIRES FOR HYBRIDIZATION MONITORING

Highly sensitive protocols for monitoring DNA

hybridization or single viruses in connection to ptype silicon NW (SiNW) functionalized with PNA probes or antibodies for influenza demonstrated [10]
Discrete conductance changes, characteristic of the

binding event observed at extremely low target concentrations.

[10] Nano Lett. 4, 5155.

DNA hybridization measurements at SiNW functionalized with PNA probes. (A) Real-time conductance response to 60 fM WT DNA sample. The arrow marks the point in time when the sample was added. The inset shows a SEM image of a typical SiNW device with source (S) and a drain (D) indicated; scale bar is 1m. (B) Time dependent conductance in DNA-free solution; the arrow indicates the point in time when a new solution sample was added [13]

[13] Biosens. Bioelectron. 2006, 21, 1887-1892.

NANOPARTICLES FOR CANCER MARKER DETECTION

Ultrahigh sensitivity of nanoparticle-based

electrochemical sensing protocols: opens up possibility of detecting cancer markers that cannot be measured by conventional methods. Stripping voltammetry: useful for detecting metal nanoparticle tags due to accumulation (electrodeposition) step. Use of gold nanoparticle tracers for stripping-based electrochemical detection of DNA hybridization and antibodyantigen interactions reported [11] Relied on capturing the gold nanoparticles to hybridized target or captured antigen, followed by dissolution and electrochemical stripping measurement of metal tracer
[11] J. Pharm. Biomed. Anal. 19, 5374

INORGANIC NANOCRYSTALS FOR PROTEIN DETECTION

Inorganic nanocrystals offer electrodiverse population of

electrical tags as needed for multiplexed clinical testing. Use of different inorganic-nanocrystal tracers for multitarget electronic detection of proteins reported [12] Four encoding nanoparticles (cadmium sulfide, zinc sulfide, copper sulfide, and lead sulfide) used to differentiate signals of four proteins or DNA targets in connection with sandwich immunoassay along with stripping voltammetry of corresponding metals Each binding event yielded distinct voltammetric peak, whose size and position reflected the level and identity, of the corresponding antigen or DNA target
[12] J. Am. Chem. Soc., 2003 125, 32143215.

Simultaneous monitoring of multiple proteins in connection to different inorganic nanocrystal tags and electrochemical stripping transduction [14]

[14] Anal. Chem. , 2003, 81, 71217129.

CONCLUSION
One dimensional nanostructures: widely attractive for wide

range of bioelectronic sensing applications. The ability to modify nanowires and nanotubes with biological recognition elements- high selectivity. Successful utility of 1-D nanostructures requires new nanofabrication capabilities with proper interconnection, reproducible positioning of nanowires and nanotubes between closely-spaced microelectrodes. Proper attention to be given to interface of these devices with the real world (sample delivery) Nanomaterials-based electrochemical devices- expected to have major impact upon clinical diagnostics, environmental monitoring, security surveillance, food safety.