Overview and Introduction to Biosensors

Anthony P.F. Turner

Biosensors and Bioelectronics Centre

anthony.turner@liu.se www.ifm.liu.se/biosensors

TFAY62 - Tuesday 27 January 2015

Overview and Introduction to Biosensors

• What is a biosensor?

• Why are biosensors important?

• Electrochemical biosensors

• Affinity biosensors

• Whole-cell biosensors

• Emerging technologies

• Conclusions & further reading

2

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 applicationsin:

• 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!!!

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“A biosensor is an analytical device incorporating a biological orbiologically 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 (Elsevier)

The Biosensor – a definition

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

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, biomimetic 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

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

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

• Magnetic: e.g. magneto-resistive devices, paramagnetic labels etc

• Micromechanical: e.g. resonating beam structures

Types of Biosensor

Catalytic Biosensore.g. enzyme electrode

Labelled Affinity Sensore.g. Fluorescence or Enzyme labelledImmunosensor

Label-free Affinity Sensor e.g. SPR, piezoelectricor electrochem

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Scopus “biosensor*”

2 papers on Biosensors identified in 1980, 75 papers in 1985 and ~5,000 last year

Biosensor Publications

>59,000 papers on Biosensors

Biosensors: World

Market

80% of market is still for glucose, but we are on the cusp of a new era

World Market for Biosensors (US$m)

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

02000400060008000

1000012000140001600018000

1996 2000 2002 2004 2006 2009 2010 2018

Biosensors: $13b Market Share

Bayer Contour

Leaders in the US$13 billion marketBlood GlucoseRoche DiagnosticsJ&J LifescanBayer DiagnosticsAbbottBeijing Yicheng JPS-5Subcutaneous GlucoseMedtronicDexcomOthersMolecular Devices Corp (LAPs Affinity)Affymetrix etc (DNA chip)Nova Biomedical etc (Critical care)BIAcore etc (SPR affinity)

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Some key drivers

Healthcare spending is growing unsustainably: 18% of GDP for USA, 9.6% of GDP for Sweden

• Individual choice and ownership of data

• Consumer-driven delivery with evidence-based reimbursement

• Decentralisation and radical restructuring of services

• Personalised Medicine

• Mobility

As much data as all of humanity as a whole created from the dawn of civilisation until 2013, is now being created every other day.

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In-Body Sensors: a Trillion US$ Healthcare Opportunity

Gate & speed Head impact (sports)Heart, skin, breathing Exposure to sunBody temperature Biomechanical dataCalories & distance Altitude & rate ascentSleep patterns Location (3D)Brainwaves & control Speed & accelerationPosture Repetitive activity

“There is an obvious omission from the above list: there is presently no way to measure aspects of blood chemistry, or other parameters that can only measured by sensor technology that reside inside the body.”

Wearable Technology: 2014. Company Profiles, Market Analysis & Forecasts. www.generatorresearch.com

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

2

2015

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Mediated Amperometric Glucose Sensors

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 amperometricdetermination of glucose. Analytical Chemistry 56, 667-671.

MediSense ExacTech™ 1987

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Driving down cost – Screen printing

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The first pocket-sized device

• The creation of the 1st pocket-sized electrochemicalanalyser for home blood glucose monitoring in 1987 laidthe foundations for the current generation of instruments (e.g. Abbott, Roche, J&J and Bayer)

• US$10,000+ analogue lab instruments had to be reducedto programmable devices, eventually costing $7-15

• Production of hand-made enzyme electrodes had to be automated (current costs 2-5.5 cents)

• Biochemistries had to be formulated

as bioprintable materials

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

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 iPhoneplug-in glucose meter gainedFDA 510(k) on 7 December 2011, marketted by Sanofi Aventis with iBGStar app,2012.

Short Break

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The Arrival of Continuous Glucose Monitoring (CGM)

Medtronic Dexcom Abbott FreestyleGuardian STS Navigator

Meter Kit $1,339 $800 $960-1,040Sensors/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 16Na+, K+, Cl-, TCO2, Glu, BUN (urea), Creatinine& Hct

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GlucoseCreatinineLactateUrea

SodiumCalciumPotassiumChloridepHpCO2Haematocrit

i-Stat Clinical Analyser

1992

Surface Plasmon Resonance (SPR) - BIAcore

Ingemar, Claes & Bo, LiU

Bo Liedberg, Claes Nylander and Ingemar Lunström (1983)Surface plasmon resonance for gas detection and biosensingSensors 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

Biosensors Applications AB: technology for drug and explosives detection

Biosens 600

www.biosensor.se

Border controlSecurity Correctional & Police authorities Rehabilitation & Workplace control

Microbial Growth Detection with Resonating BeamsResonating 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 now routinely used for imaging genomic and proteomic arrays

Multianalyte detection based on total internal reflection on planar

waveguidesZeptosens (Bayer)

Raptorwww.resrchintl.com

Lateral Flow Affinity Assays

Membrane coating Membrane drying and cutting

Lateral flow device assembly

Dispenser Membrane cutter Assembly roller

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 hCGusing 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 pregnant2) Pregnant with conception 1-2 weeks ago3) Pregnant with conception 2-3 weeks ago4) 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.

AB

Whole-cell biosensors: Biochemical Oxygen Demand (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).

NanomaterialsNanomaterials have impacted enormously on biosensor R&D over the last 10 years and are expect to filter through to commercial products in the near future

Quantum Dots

Molecularly-imprinted Polymers

Nanowires

Carbon Nanotube (Graphene) with dendrimer

Aptasensors

AB

Thrombin aptamer (TBA) 5’-GGTTGGTGTGGTTGG-3’ interacts specifically with the Fibrinogen recognition exosite through the two TT loops

Bini, A., Mascini, M., Mascini, M. and Turner, A.P.F. (2011). Selection of Thrombin-binding aptamersby 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.

Molecular Imprinting (Plastic antibodies)

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.

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Printed Electronics Arena at Norrköping

Dry Phase Patterning- Metallic (Al) foil laminates- Patterning through dry process- Up to 150 m/min- Web width: 300 mm- Environmentally friendly

DimatixInkjet- 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

NILPETERRoll-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

2 Flat bed screen printer s & 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

Hybrid line to be added shortly

Towards the fully-printable instrument

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

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!

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

www.ifm.liu.se/biosensors

6.4512013

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Some Web Siteswww.ysilifesciences.com YSI Analyserswww.minimed.com/products/guardian Implantable glucose sensorswww.youtube.com/watch?v=GxThyTTztmQ video: Printing sensorswww.accu-chek.com.au/au/products/metersystems/advantage.htmlwww.bayerdiabetes.com/sections/ourproducts/meters/breeze2www.lifescan.com/ www.abbottdiabetescare.com/index.htm www.biacore.com/lifesciences/index.htmlwww.ifm.liu.se/biosensors