Overview and Introduction to Biosensors Anthony P F Turner

Biosensors & Bioelectronics Centre IFM-Linköping University

Biosensor Technology TFYA62, Linköping, , Linköping, Tuesday 28 January 2014

Biosensors & Bioelectronics Centre

The Centre’s mission is to harness the fundamental research activities and innovation at LiU to facilitate the creation of the next generation of bioelectronic devices and to support the national and worldwide development of the field. Activities encompass:

Fully Integrated Sensors

Non-invasive Sensing

Wearable and Distributed

Sensors

www.ifm.liu.se/biosensors

Anthony P F Turner

• Originally a Biochemist, since 1980 helped establish the field of Biosensors: • Edited the principal journal in the field, Biosensors & Bioelectronics, since co-founding it in 1985 and published the first text book on Biosensors in 1987 • Founded the World Congress on Biosensors in 1990 and Chaired it since then • >750 publications and patents in the field of biosensors and biomimetic Sensors; >400 keynote and plenary lectures; H Index 63

• 35 yr academic career in the UK culminated in the positions of Principal of Cranfield University at Silsoe and Distinguished Professor of Biotechnology

• 2010 established a new Centre for Biosensors and Bioelectronics at LiU • Fellow of the Royal Society of Chemistry, higher doctorates for exceptional contribution to biosensors and for contribution to higher education, and Foreign Associate of the USA National Academy of Engineering, Member of the Royal Swedish Academy of Engineering (IVA)

• Led the team that pioneered the technology that now dominates the home blood glucose monitoring market and continues to work for and advise companies and governments worldwide in analytical biotechnology

• Remain a Director of Cranfield Ventures Ltd, with responsibility for leveraging Cranfield University's IP via spin outs and licensing

• Served as an Expert Witness in patent litigations on three continents

Contents

• What is a biosensor?

• Why are biosensors important?

• Electrochemical biosensors

• Affinity biosensors

• Whole-cell biosensors

• Emerging technologies

• Conclusions & further reading

The Atlas moth can follow a trail of pheromones from severl km downwind

Applications

Biosensors harness the immensely powerful molecular recognition properties of living systems and engineer these into electronic devices to provide easy-to-use sensing devices with applications in: – Medicine – Biomedical research – Drug discovery – Environmental monitoring – Food content, quality and safety – Process control – Security and defence

The two most successful biosensors to date: • Mediated amperometric glucose biosensor • Real-time bioaffinity interaction analysis

Newman, J.D. and Turner, A.P.F. (2005) Home blood glucose biosensors: a commercial perspective. Biosensors and Bioelectronics 20, 2435-2453.

The Biosensor

Bioreceptor

Transducer

GAS!!!

Economic Impact • Healthcare spending is growing fast: 18% of GDP for USA, 9.5% of GDP for Europe

• Global Healthcare spending is more than 5 Trillion Dollars per year

• This spending trend is unsustainable for the future economy

• So what does this mean for the future?

• Individual choice and ownership of data • Consumer-driven, outcomes-based delivery • A boom in Personalised Medicine • Decentralisation and radical restructuring of

services • Evidence-based reimbursement • MOBILITY !!!

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Scopus “biosensor*”

This simple search shows 1 paper on Biosensors identified in 1980, 75 papers in 1985 and 4,819 in 2013

Biosensor Publications

~10% of all papers ever published on biosensors

appeared in 2013

Biosensors: World

Market

85% of Biosensor market is still for glucose measurement

World Market for Biosensors (US$m)

Turner, A.P.F. (2013) Biosensors: sense and sensibility. Chemical Society Reviews 42 (8), 3184-3196.

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1000012000140001600018000

1996 2000 2002 2004 2006 2009 2010 2018

Roche Diagnostics

Lifescan

Bayer Diagnostics

Abbott

Molecular Devices Corporation

Affymetrix

Nova biomedical

Biacore

Others

China

Biosensors: $13b Market

Share

Beijing Yicheng JPS-5

Roche Accu-Check Aviva Nano

Lifescan OneTouch Ultra

Bayer Contour

Abbott FreeStyle Lite

The Market leaders in Biosensor Sales

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“A biosensor is an analytical device incorporating a biological or biologically derived sensing element either intimately associated with or integrated within a physicochemical transducer. The usual aim is to produce a digital electronic signal which is proportional to the concentration of a chemical or set of chemicals.” “Biosensors usually yield a digital electronic signal which is proportional to the concentration of a specific analyte or group of analytes. While the signal may in principle be continuous, devices can be configured to yield single measurements to meet specific market requirements.” (One-shot biosensors) Turner, A.P.F., Karube, I. and Wilson, G.S. (1987). Biosensors: Fundamentals and Applications. Oxford University Press, Oxford. 770p. ISBN: 0198547242 & The international journal Biosensors & Bioelectronics

Biosensors

Receptors for Biosensors

• Biological materials: e.g. tissue, microorganisms, organelles, cell receptors, enzymes, antibodies, nucleic acids, natural products etc

• Biologically derived materials: (e.g. recombinant antibodies, engineered proteins, aptamers etc

• Biomimics: e.g. synthetic receptors, bimimetic catalysts, combinatorial ligands, imprinted polymers etc

Transducers for Biosensors

• Electrochemical: e.g. amperometric, potentiometric, conductimetric, impedimetric etc

• Optical: e.g. surface plasmon resonance (SPR), fluorescence, interferometric, holographic

• Thermometric: e.g. enzyme thermistor, thremal enzyme-linked immunosorbent assay etc

• Piezoelectric: e.g. quartz crystal microbalance (QCM), surface acoustic wave devices (SAW)

• Magnetic: e.g. magneto resistive devices, paramagnetic labels etc • Micromechanical: e.g. resonating beam structures

The Biosensor

(Bio)receptor(affinity or catalytic)

TransducerElectronics

Electrochemical

Optical

Thermometric

Piezoelectric

Magnetic

Micromechanical

Electrochemical

Optical

Thermometric

Piezoelectric

Magnetic

Micromechanical signal

Analytes

TissuesMicroorganismsOrganellesCell receptors

EnzymesAntibodiesNucleic acidsSynthetic receptors

The Biosensor

Types of Biosensor

Catayltic Biosensor e.g.enzyme electrode

Labeled Affinity Sensor e.g. Fluorescence or Enzyme labeled Immunosensor

Label-free or direct Immunosensors e.g. SPR or piezoelectric

The Starting Point: Clark Oxygen Electrodes

1955

Yellow Springs Instrument Company Inc (YSI)

Glucose Biosensor 1975

YSI, Ohio 1987

The original YSI serum-glucose biosensor for diabetes clinics 1975

Clark, LC & Lyons, C (1962). Annals New York Academy of Sciences 102, 29.

1987

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Enzyme Electrochemistry

Cass, A.E.G., Davis, G., Francis, G.D., Hill, H.A.O., Aston, W.J., Higgins, I.J., Plotkin, E.V., Scott, L.D.L. and Turner, A.P.F. (1984) Ferrocene-mediated enzyme electrode for amperometric determination of glucose. Analytical Chemistry 56, 667-671.

Glucose oxidase or PQQ Glucose Dehydrogenase

Mass Production: Screen Printing

• Bayer’s DIDGET™ blood glucose meter plugs into a Nintendo DS™ or Nintendo DS™ Lite system

• This helps encourage consistent testing with reward points that children can use to buy items and unlock new game levels

The Importance of the User Interface

Blood Glucose: “We’ve got an App For That”

Lifescan popularised the iPhone route 2009

AgaMatrix Nugget iPhone

plug-in glucose meter gained FDA 510(k) on 7 December 2011, to be marketted by Sanofi Aventis with iBGStar app, early 2012.

The Arrival of Continuous Glucose Monitoring (CGM)

Medtronic Dexcom Abbott Freestyle Guardian STS Navigator Meter Kit $1,339 $800 $960-1,040 Sensors/m $350 (10x3day) $240 (4x7day) $360-390 (6x5 day) FDA Aug 2005 March 2006 March 2008 (CE June 07) approval Reading 1 per 5min (2h run in) 1 per 5min (2h) 1 per min (10h run in) Frequency Reading must be checked by finger-stick method before adjusting insulin

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Nova Biomedical CRT 16 Na+, K+, Cl-, TCO2, Glu, BUN (urea), Creatinine & Hct

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Glucose Creatinine Lactate Urea Sodium Calcium Potassium Chloride pH pCO2 Haematocrit

i-Stat Clinical Analyser

1992

Short Break

Surface Plasmon Resonance (SPR) - BIAcore

Ingemar, Claes & Bo, LiU

Bo Liedberg, Claes Nylander and Ingemar Lunström (1983) Surface plasmon resonance for gas detection and biosensing Sensors and Actuators 4, 299-304

Chemosensing based on angle-resolved surface plasmon resonance is demonstrated on intact cell phones using a disposable optical coupler and software to configure illumination and acquisition. This coupler operates on different cell phones and is applied for classical affinity assays with commercial chips and custom-made tests with embedded calibration. Measured performance (2.14x10−6 refractive index units) is comparable with compact SPR systems.

Preechaburana, P., Gonzalez, MC., Suska, A., Daniel Filippini, A. (2012). Surface Plasmon Resonance Chemical Sensing on Cell Phones. Angewandte Chemie 51, 11585–11588.

SPR interfaced to a Mobile Phone

SPR Imaging Aptasensors

Scarano, S, Mascini, M., Turner, A.P.F. and Minunni, M (2010) Surface Plasmon Resonance Imaging for Affinity-Based Biosensors. Biosensors & Bioelectronics 25, 957-966

Kretschmann configuration for SPRi; p-polarised light is directed to the prism, on which the biomolecular probe is tethered, and a CCD camera collects the output signal as variations in reflectivity

Horiba – up to 900 spots Light activated peptide synthesis 2,500 spots; target 7,500 for epitope mapping etc

Biosensors Applications AB: technology for drug and explosives detection

Biosens 600

www.biosensor.se

Border control Security Correctional & Police authorities Rehabilitation & Workplace control

Microbial Growth Detection with Resonating Beams Resonating Beam Structures for Antibiotic Testing

Fluoresence-based affinity sensors

Up to 6.5 m assays on a

1.3 cm2 chip Fluorescence immunoassay-based biosensor for real time or near real time detection of microbial pathogens, typical

assay times of 10-15 minutes; 4 disposable optical

waveguide sensors; may be reused if test results continue

to be negative.

DNA Chip

Confocal scanners and other microscopy techniques are now routinely used for imaging genomic and proteomic arrays

Leading methods for multianalyte detection based on total internal reflection on planar waveguides

Zeptosens (Bayer)

Raptor www.resrchintl.com

Lateral Flow Immunoassay

Original visually read pregnancy test format patented by Unipath and launched in 1988, based on blue latex label resulting in one or two blue lines indicating hCG

Pregnancy Tests with Conception Indicator Launched July 2008, measures urinary hCG using both a low sensitivity and a high sensitivity strip. Control lines and results read using red light to measure density of lines by reflectometry yielding 4 possible semi-quantitative results: 1) Not pregnant 2) Pregnant with conception 1-2 weeks ago 3) Pregnant with conception 2-3 weeks ago 4) Pregnant with conception >3 weeks ago

Swiss Precision Diagnostics GmbH is a joint venture between Procter and Gamble and Inverness Medical

>99% accurate in detecting pregnancy & 92% accurate in time predictions. Same price as visual strips.

A

B

Whole-cell biosensors: BOD

Whole-cell Biosensors • Microorganisms engineered to react to the presence of

chemical signals with the production of an easily quantifiable marker protein

• The regulatory system in the bacterial cell is exploited to drive expression of a specific reporter gene, such as bacterial luciferase, green fluorescent protein, beta-galactosidase

• This is achieved by fusing the DNA for a promoterless reporter gene to an extra copy of the selected regulatable promoter and introducing this construction into the bacterial or yeast cell

• Examples include heavy metal resistance (heavy metal sensors), organic compound degradation (organic compound sensors), cellular stress responses (to obtain general toxicity sensors) and DNA damage repair (mutagenicity).

Nanotechnology Materials The market for pure nanomaterials (carbon nanotubes, nanoparticles, quantum

dots, dendrimers etc.) was about $3.6 billion by 2010 (Lux Research). Many of these materials will find application in diagnostics

Aptasensors

A

B

Bini, A., Mascini, M., Mascini, M. and Turner, A.P.F. (2011). Selection of Thrombin-binding aptamers by using computational approach for aptasensor application. Biosensors and Bioelectronics 26, 4411-4416.

SELEX searches 1014 of possible 1018 20-60mers

Poma, A., Turner, A.P.F. and Piletsky, S. (2010) Advances in the manufacture of MIP nanoparticles. Trends in Biotechnology 28, 629-637.

Synthetic Receptors e.g. Molecular Imprinting

Reversible interactions between the template and the polymerisable functional monomer may involve: (a) reversible covalent bonds, (b) covalently attached polymerisable binding groups that are activated for non-covalent interaction by template cleavage, (c) electrostatic interactions, (d) hydrophobic or van der Waals interactions. Following polymerisation, the template is then removed through the disruption of the interactions with the polymer, and subsequently extracted from the matrix. The target analyte or his analogues can then be selectively rebound by the polymer.

Nano-MIPs (Plastic Antibodies) a)

c)

b)

d)

e)

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AB A + B. .λν

.A + B. A B

+ M .A . A

BA λν .+ BA .

Karim, K., Piletsky, S.A., Piletska, E.V., Turner, A.P.F. Chianella, I. and Guerreiro, A. Preparation of soluble and colloidal molecularly imprinted polymers by living polymerization. US Patent 8,192,762 granted 5 June 2012

Switchable Immunosensors

Deshpande, S., Tiwari, A. and Turner, A.P.F. (2012). Label-free nano immunosensor for diagnosis of cardiac injury based on localised surface plasmon resonance. Label-free Technologies, 1-3 November 2012, Amsterdam, Elsevier.

Poly(N-isopropylacrylamide) {PNIPAAm} temperature-responsive polymer at >32°C undergoes a reversible lower critical solution temperature phase transition (LCST) from swollen hydrated state to shrunken dehydrated state, losing ~90% of mass

Gold nanorod-based plasmonic biosensing displayed much higher selectivity and sensitivity in the picograms/ml range for trace analytes

Putting the “Bio” into Printed Electronics

Dry Phase Patterning - Metallic (Al) foil laminates - Patterning through dry

process - Up to 150 m/min - Web width: 300 mm - Environmentally friendly

Dimatix Inkjet - Bioprinting: DNA, lipids, proteins - Ag ink, PEDOT:PSS ink - 1-10 pL drops - Min ink amount to be printed: 0.5 ml - Substrate thickness 40 µm-25 mm - Substrate heating up to 60 °C

NILPETER Roll-to-roll label printer for screen & flexo - 5 printing stations (flexo/screen) - Dryers (hot air/UV curing)

- Web width: 180-330 mm - Web velocity: speed 3-180 m/min - Die cutting - Lamination - Min feature size: 100 µm - Flexible substrates

Flat bed screen printer & Conveyor feed dryer - Rigid/flexible substrates - Vacuum substrate table - Substrate size <DIN A6-DIN A3 - Pneumatic driven filler and squeegee - Registration accuracy ~50 µm - Minimum feature size: 100 µm - Min ink 150 ml/printing unit (screen) - Hot air/UV/IR drying units

1 Roll-to-roll label printer (flexo, screen) 2 Flat-bed screen printers 1 Conveyor feed dryer 2 UV dryers 2 Ink Jet printers 1 Dry Phase Roll-to-roll patterning machine

• Kliche making • Inspection and quality control • Environmental testing • Ink development and manufacturing • Design

Printable components developed by Acreo, Linköping University (and partners)

Monochrome Emissive or reflective Paper or plastic 1-3 V (reflective) 110 V (emissive)

Displays

All-printed , Mn -ZnO Capacity: 1-10 mA h (www.enfucell.com)

Enzyme electrodes Electrolyte sensors Piezo and Pyroelectrical Humidity, temperature, heat

,pressure, vibrations Electrochemically &

electrolyte gated OFET 0.5 – 1.5 V Switch time: 10-6 to 10-2 s

Non-volatile and flexible Based on ferroelectric

polymers Retention time: > 10

years 109 cycles Read/write: ms Fully qualified (www.thinfilm.se)

Metal Al, Cu 1 kHz – 1 GHz Resolution: 100 mm Material thickness: 1-10

mm Fully qualified (www.webshape.se)

All-printed on antennas/electrodes

High frequency Performing

qualification

Transistors

Sensors

Batteries

Memories Antennas

Diodes

Turner, A.P.F. (2013) Biosensors: sense and sensibility. Chemical Society Reviews 42 (8), 3184-3196.

Integrated Printed Disposable Biosensor

Turner, A.P.F. (2013) Biosensors: sense and sensibility. Chemical Society Reviews 42 (8), 3184-3196.

Printed Transistors For Display control

Silicon chip

Printed Display

Biosensor Sample Deposition

Printed Battery

Conclusions • Biosensors have achieved considerable success in both the commercial and academic arenas and the need for new, easy-to-use, home and decentralised diagnostics is greater than ever

• Amperometric biosensors continue to dominate the market

• Electrochemical biosensors are available for a range of decentralised analyses, including medical, food and environmental applications

• Array-based fluorescence sensors are now well established for genomic and proteomic assays

• Label-free assays based on SPR dominate the laboratory affinity sensor market especially in drug development

• The search continues for a new disruptive technology!

5.437 2012

www.ifm.liu.se/biosensors

Turner, A.P.F. (2013) Biosensors: sense and sensibility. Chemical Society Reviews 42 (8), 3184-3196. Newman, J.D. and Turner, A.P.F. (2008). Historical perspective of biosensor and biochip development. In: Handbook of Biosensors and Biochips (Eds R. Marks, D. Cullen, I. Karube, C. Lowe and H. Weetall) John Wiley & Sons. ISBN 978-0-470-01905-4 www.wiley.com/go/biosensors Newman, J.D. and Turner, A.P.F. (2005). Home blood glucose biosensors: a commercial perspective. Biosensors and Bioelectronics 20, 2435-2453

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Some Web Sites

www.ysilifesciences.com www.minimed.com/products/guardian www.youtube.com/watch?v=GxThyTTztmQ (video) www.accu-chek.com.au/au/products/metersystems/advantage.html www.bayerdiabetes.com/sections/ourproducts/meters/breeze2 www.lifescan.com/ www.abbottdiabetescare.com/index.htm www.biacore.com/lifesciences/index.html www.ifm.liu.se/biosensors