Countries Afgected by Landmines Countries afgected by Landmines - - PowerPoint PPT Presentation

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Countries Afgected by Landmines Countries afgected by Landmines - - PowerPoint PPT Presentation

Jul 17, 2023 608 likes •1.02k views

e ectrace Countries Afgected by Landmines Countries afgected by Landmines Image adapted from Handicap International Afgected Countries Non Afgected Countries 2 Conceptual Design Wifj enabled device reads Electrace current and sends data to

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e ectrace

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Countries Afgected by Landmines

Countries afgected by Landmines Image adapted from Handicap International Afgected Countries Non Afgected Countries

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Conceptual Design

μtrace contains E. coli (Landmine) biosensor

e

Read-out from Electrace Wifj enabled device reads Electrace current and sends data to web app.

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Biosensor

  • Landmines

Outside the Lab

  • Microfmuidic Platform

Electrical Signal

  • Conductive Curli
  • Electron Transport

INPUT OUTPUT μTRACE

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Biosensor

  • Landmines

Outside the Lab

  • Microfmuidic Platform

Electrical Signal

  • Conductive Curli
  • Electron Transport

INPUT OUTPUT μTRACE

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Improving the Response

BioBrick (BBa_K1316008)

Literature: Yagur-Kroll et al., 2014

Time (hr)

100mg/L DNT 50mg/L DNT 0mg/L DNT 100mg/L DNT +N 50mg/L DNT +N 0mg/L DNT +N

ybiJ Promoter

5 5 10 15 5 10 15 20 25

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Biosensor

  • Landmines

Outside the Lab

  • Microfmuidic Platform

Electrical Signal

  • Conductive Curli
  • Electron Transport

INPUT OUTPUT μTRACE

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Biosensor

  • Landmines

Outside the Lab

  • Microfmuidic Platform

Electrical Signal

  • Conductive Curli
  • Electron Transport

INPUT OUTPUT μTRACE

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Curli in Biofjlm

BioBrick (BBa_K1316014)

csgB csgA Constitutive Inducible B A A A B A A A B A A A A A A A A

Induction

Cell Membrane

Barnhart and Chapman, 2006

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Conductive Curli

Cell with Curli Cell without Curli Gold nanoparticle Medium Gold Electrode Substrate

Induction

Chen et al., 2014

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Stochastic Curli Modeling

Gene Level

nm scale

Cell Level

μm scale

Colony Level

mm scale

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Stochastic Curli Modeling

Deterministic

ODEs

Stochastic

Worm Like Chains Percolation Theory & Graph Theory

Stochastic

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Percolation

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Percolation

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Conclusions from Modeling

Induction of conductive Curli results in an electrical signal. This is a qualitative response.

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e- Biosensor

  • Landmines

Outside the Lab

  • Microfmuidic Platform

Electrical Signal

  • Conductive Curli
  • Electron Transport

INPUT OUTPUT μTRACE

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Shewanella oneidensis MR-1

Adapted From Goldbeck et al., 2013

Extracellular Space Cytosol Lactate Dehydrogenase Q QH2 MtrB MtrC MtrA CymA Electron Donor Electron Donor Oxidized Metal Oxides

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Escherichia coli C43

The fjrst functioning MtrCAB Cluster (BBa_K1316012)

Electrode Extracellular Space Cytosol Lactate Dehydrogenase Q QH2 Electron Donor Electron Donor Oxidized MtrB MtrC MtrA NapC

Cytochrome C Maturation System (BBa_K1316011)

Adapted From Goldbeck et al., 2013
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Flux Balance Analysis

Electrode Extracellular Space Cytosol Lactate Dehydrogenase Q QH2 Lactate Pyruvate MtrB MtrC MtrA NapC Anoxic

Determination of Optimal Electron Donor Using COBRA Toolbox

Adapted From Goldbeck et al., 2013
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Electron Flow

A

Donor Oxidized Donor e- e- e- e- e- Working Electrode Counter Electrode Reference Electrode MtrCAB

+

Potentiostat

  • e-
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Custom-designed Bioreactor

Magnetic stirrer-bar Heating mantle Carbon cloth working electrode Nitrogen inlet Sampling tube Gas outlet Silver/Silver Chloride reference electrode Counter Electrode

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Custom-designed Bioreactor

Software Development in LabVIEW

Experimental Setup Potentiostat Circuit and Software

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Measurement of Current

  • E. coli C43 +

MtrCAB & Ccm

  • E. coli C43

Normalized Currnet [mA] 10 9 8 7 6 5 4 3 2 1 Time [hr] 4 3 2 1

A e- e- e- e- e-

+

  • MtrCAB & Ccm BioBrick (BBa_K1316012 & BBa_K1316011)
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Biosensor

  • Landmines

Outside the Lab

  • Microfmuidic Platform

Electrical Signal

  • Conductive Curli
  • Electron Transport

INPUT OUTPUT μTRACE

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Microbial Biosensors

Now: Numerous but Impractical

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Microbial Biosensors

Now: Numerous but Impractical Our System: Cheap, Easy to handle for non- biologists in the fjeld, plug-&-play, quantifjable

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Microbial Biosensors

“Alternative read-outs are very interesting. For a device like yours, which is able to be applied in the fjeld, this is a really nice trick.” - Dr Van Der Burg (BioDetection Systems) Now: Numerous but Impractical Our System: Cheap, Easy to handle for non- biologists in the fjeld, plug-&-play, quantifjable

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B t By ofgering a synbio solution to solve an every day problem, we prove to the broader public that they need synthetic biology in their life.

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“If your system is capable of detecting land mines, I wouldn’t mind if it uses synthetic biology.” - Suzanne Oosterwijk, PAX By ofgering a synbio solution to solve an every day problem, we prove to the proader public that they need synthetic biology in their life.

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μtrace - a Custom-designed Device

Fluid Channel Commercial Electrode Glass Slide Electrical Contacts PDMS 33 μL of Analyte

Inexpensive Prototyping

~$3

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μtrace - a Custom-designed Device

Inexpensive Prototyping

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Potentiostat μtrace Infmow Outfmow Working Electrode Counter Electrode Reference Electrode

A

Syringe Pump

Experimental Setup

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μtrace test results - Ascorbic Acid Voltammetry

0.4 0.6 0.8 1.0 1.2 1.4 1.6 0.2 1.8 Ascorbic Acid 0.2V Ascorbic Acid 0.4V Water 0.4V Water 0.2V

Time (s) 50 100 150 200

ybiJ Promoter

250 Current (mA) 300

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μtrace with MtrCAB & Ccm

0.005 0.01 0.015 0.02 0.025 0.03 0.035 0.04 0.045

Current (μA)

  • E. coli C43 + MtrCAB & Ccm
  • E. coli C43

Water

Time (s) 50 100 150 200 250 300

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Achievements

Improved responsiveness of modifjed E. coli to detect landmines Stochastic modeling of Curli shows a qualitative response Improved the formerly registered MtrCAB BioBrick Modifjed E. coli to produce an electrical output Designed μtrace - a microfmuidic device for measurement

  • f electrical output
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e ectrace

A plug-&-play platform for biosensors, that produces a quantitative electrical output that can be used for the detection of landmines.

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Partner:

TU Delft Department

  • f Biotechnology

TU Delft Faculty of Applied Sciences

Sponsors:

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e ectrace

A plug-&-play platform for biosensors, that produces a quantitative electrical output that can be used for the detection of landmines.