Zurich, April 27th, 2012

G. De Micheli, Q. Huang, L. Thoeny-Meyer, Y. Leblebici, C. Dehollain, F. Grassi, S. Carrara

Implantable/Wearable System for on-line Monitoring of Human Metabolic Conditions

(Implantable-IRONIC)

Outline

Introduction:
Objectives, motivation and roadmap

New nano-biosensor technology

Nanostructured sensors Experimental results and comparisons

New micro-electronic circuits

Data acquisition and energy harvesting System bio-compatibility and tests

Conclusions and outlook

2

Project objective

Remotely-powered cylinder Dimension: 2.5x15mm

Design implantable/wearable systems for continuous monitoring of human metabolism

Engineering roadmap

November 2010
I1: Prototype of glucose sensor (10x32mm)

November 2011
I2: Prototype of sensor for various metabolites and experiments with animals (10x32mm) I3: Integrated components for sensor (2.2x15mm)

Spring 2013
I3: Assembly and test of multi-metabolite sensor (2.2x15mm)

Prototype I2
On-mouse telemetry system

Patch-model Implanted nano-bio-sensor targeting various metabolites

Collaboration with antenna group

4.8 mm

7.8mm

13.1mm

24.7mm

Prototype I3

Prototype I3
IC Integrated CircuitRF and detection CMOS circuit for
Temperature Sensor 2.2 mm pH Sensor Molecular Sensors

inductive coil

Components for new prototype I3

Passive multi-sensor silicon interposer (2.2x12mm) Data acquisition chip (1.5x1.5mm) Micro-antenna(2x15 mm) Enclosure (not shown)

External data/power patch

Features
Remote powering of implantable biosensors through inductive link Short-range bidirectional communication with the implanted sensors Long-range communication with remote devices

Advantages
Improved wearability Possibility to place it directly over the implant area Completely stand-alone, no wires are needed Battery powered

Scientific highlights

Realization and test of nano-structured sensors for various metabolites

Increased sensitivity and lower LOD New genetically-engineered probes for higher robustness

Integrated low-power programmable data acquisition electronics Power and data transmission means

Multi-layer inductive coil for power/data transmission Experimentation with various means of transmission
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Outline

Introduction:
Objectives, motivation and roadmap

New nano-biosensor technology

Nanostructured sensors Experimental results and comparisons

New micro-electronic circuits

Data acquisition and energy harvesting System bio-compatibility and tests

Conclusions and outlook

12

Sensing various metabolites

TARGETS

Probe Enzymes
Glucose Oxidase Lactate Oxidase Glucose Oxidase & Hexokinase

Endogenous metabolites
Glucose Lactate ATP

Glutamate Oxidase Glutamate

Nano-Bio-Sensors Macro-Assembly
BARE ELECTRODE

CARBON NANOTUBES

10.3 1.14 nm
CNTs + PROBE ENZYMES

19.9 3.38 nm 4.9 nm

3.6 nm
Boero, Carrara et al. / IEEE PRIME 2009 Boero, Carrara et al. / IEEE ICME 2010 De Venuto, al. et Carrara / IEEE Senors 2010 Boero, Carrara et al. / Sensors & Actuators B 2011 Carrara et al. / Biosensors and Bioelectronics 2011 Boero, Carrara et al. / IEEE T on NanoBioScience 2011

5.2 nm

Selective electro-deposition of probes

CHITOSAN
Chitosan Polysaccharide Liquid-to-solid state according to pH Electric field to: Put probe in position and alter pH Entrap enzymes and CNT Biocompatible and reversible

Array 40

Array 10

Array 10 bright field

Array 10 dark field

Electrodeposition
600 +1.5 V Chitosan 0.7% CNT 1 mg/ml pH 5

Electrocleaning
600 -2V PBS 1x pH 7.4

In-vitro measurement setup

Base Station

PC

Implant Cable

Electrode capsule

L. Bolomey 16

Glucose Monitoring

Continuous monitoring of glucose with the single metabolite remote system and a glucose Bio-Nano-Sensor

Lactate Monitoring

Continuous monitoring of lactate with the single metabolite remote system and a lactate nano-biosensor

Increased sensitivity

~ 7.5 times more

Sensor sensitivity is enhanced by nano-structuring the electrodes

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System Sensitivities System Sensitivities

Metabolite Glucose Lactate Glutamate ATP Sensitivity 27.7 A/mM cm2 40.1 A/mM cm2 25.5 A/mM cm2 3.42 A/mM cm2 Range 0.5 4 mM 0.5 2.5 mM 0.5 2 mM 0.5 1.4 mM Limit of Detection (S/N = 3) 73 M 28 M 195 M 208 M

Sensitivity and range of the bio-nano-sensors

Engineered enzymes for biosensors

Motivation: fabrication of effective and more stable biosensors for accurate diagnosis Solution: integration into biosensing platforms of tailor-designed biorecognition molecules with

higher affinity with analytes higher stability higher electron transfer rates residues able to provide an oriented or more stable immobilization
Modified enzyme a N-terminal residue

Wild type enzyme

Comparison of three Lactate Oxidases from Aerococcus viridans for biosensing

The best sensing parameters

Lactate oxidase
Active up 50 days after deposition HistidineTag Wild type Commercial

Sensitivity [A mM-1 cm-2]

35.6 6.2 18.8 6.8 26.6 5.8

Detection Limit [M]

30 6 110 21 58 21

Linear Range [mM]

0.2-1 0.2-0.8 0.2-1

40 30 20 10 0 0 20 40 Time (day) 60

Detection Limit (mM)

50 Sensitivity (A mM-1 cm-2)

200 150 100 50 0 0 20 40 Time (day) 60

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Outline

Introduction:
Objectives, motivation and roadmap

New nano-biosensor technology

Nanostructured sensors Experimental results and comparisons

New micro-electronic circuits

Data acquisition and energy harvesting System bio-compatibility and tests

Conclusions and outlook

25

VHDL-AMS cell model

Data acquisition electronics

Multi target sensing Enable CV actuation and readout Enable CA initiation and readout Low power due to remotely powering of the implantable device Low noise due to weak sensor signal

27

Circuit Design
Potentiostat and multiplexer

Biosensor current

I ()uA

Output Voltage

CA voltage (V)

Readout circuit for CV and CA readout

V (v)

Time (s)

A very slow ramp generator circuit for CV initialization

V (v)

Frontend Electronics: first prototype

Three different readout circuit blocks for CV and CA readout

The fabricated chip is compared with laboratory instruments.

Measurement results for lactate detection using the fabricated chip

Frontend Electronics: second prototype

New features:
5 different probes Enable CV actuation and readout for up to 3 targets with sub A current Enable CA initiation and readout for up to 2 targets with sub A current Embedded pH and temperature sensing that are highly needed for data calibration Low power due to remotely powering of the implantable device Low noise due to weak sensor signal Ready for system integration due to multiplexing scheme and pitch size

Temperature sensing

PH sensing

Ramp generator and CV readout

Potentiostat CA readout

IronIC Patch
Performance

Up to 15 mW transmitted within 6 mm in air

Downlink communication up to 100 kbps

Uplink communication with real-time threshold check up to 66.6 kbps

Bluetooth communication (Class-2)

Autonomy

Stand-by mode: 10 hours Power mode: 1.5 hours

Multilayer Inductors
Advantages
Inductors are realized on different PCBs, stacked, and electrically connected Link efficiency can be preserved despite area reduction

Measures on the Designed Inductors

1.17 mW (17mm Bovine Tissue) Communication is achieved at 100 kbps

Bio-compatibility study

Development of a new biocompatible polymeric package for hermetic encapsulation of individual chips at IMEC Leuven 2011-2012 Test of the biocompatible package with cell cultures to test toxicity at IMEC Leuven - 2012 Tests on induced inflammation on animal models (mice) at IRB

Tests with Animals

The Air Pouch Model in mice has been used to test the inflammatory behavior of the monitoring implants

System Biocompatibility

Tests of inflammation induced in mouse by the implanted sensor system

Outline

Introduction:
Objectives, motivation and roadmap

New nano-biosensor technology

Nanostructured sensors Experimental results and comparisons

New micro-electronic circuits

Data acquisition and energy harvesting System bio-compatibility and tests

Conclusions and outlook

37

Implantable-IRONIC Current status

Implantable/wearable system for health monitoring Top-down design:

Implant design based on four major components
Sensor, interposer, chip, antenna

External patch and communication link

Bottom-up research
Multi-target sensing

Integrated low-power electronics

Power and data transmission New target molecules

Animal models

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Implantable-IRONIC Conclusions

Contributions to bio-engineering
Design of a versatile bio-sensing platform Fusion of various disciplines Successful test of implant I2 in mice

Contributions to fundamental research

Nano-biosensors with superior properties Design of novel electronic circuits for low-power, lownoise data acquisition and tranmission New energy harvesting methods with 3D coils

Technology transfer with partner Menarini

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