ISyPeM II Giulia Cappi École polytechnique fédérale de Lausanne Annual Plenary Mee;ng of the Nano-‐Tera program May 4th, 2015
Therapeu;c drug monitoring (TDM)
Ima;nib exposure (ng/ml)
Candidates for therapeutic drug monitoring Modern anticancer agents Immunosuppressants Antiretrovirals Antibiotics
Courtesy of N. Widmer
time after dose (hrs)
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Therapeu;c Drug Monitoring Today Transfer to and measurement at central clinical labs
Interpretation of the result by a specialized doctor
Drug Concentra0on
Blood sample taken at the doctor’s office or at the hospital
Target
? Pa0ent Treatment dura0on (days)
hQp://e-‐sante.futura-‐sciences.com
AbboQ "Architect ci8200"
Today this procedure severely restricts the applicability of TDM ISyPeM II -‐ G. Cappi
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Drug quan;fica;on in body fluids Used in Clinical facilities
Agilent LC-MS/MS
Abbott "Architect ci8200” FPIA (fluorescence polarization immunoassay) CMIA (chemiluminescent microparticle immunoassay) MIA (magnetic immunoassay)
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Point-of-need
Syva RapidTest® EMIT (Enzyme multiplied immunoassay technique)
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Point-‐of-‐need therapeu;c drug Hand-held monitoring The In-Check® system integrates amplification and microarray detection processes for end-point PCR-based molecular testing in a single lab-on-chip. The In-Check system is intended to address the qualitative identification of multiple extracted nucleic-acid analytes, through PCR amplification and hybridization on a low-density microarray.
Amplification of extracted nucleic acids
Detection through low-density microarray
The integrated silicon heaters, temperature sensor and miniaturized 25 µl volume allow the In-Check PCR module to perform the rapid heating and cooling cycles required for highly reliable, end-point PCR.
The In-Check microarray module is used to detect the amplified labeled DNA sequences through hybridization on a low-density microarray composed of 126 spots.
The In-Check PCR module is thermally driven by the In-Check temperature control system (TCS). This allows fast and programmable temperature ramps together with independent random access for 5 different tests simultaneously.
iSTAT, Abbott
Accu-Chek product lines. Roche Diagnostics User-friendly operation
ISyPeM approach toTDM
The microarray module includes a set of control probes at fixed positions, as well as specific probes located according to customer requirements. The microarray module is fluidically connected to the PCR module and is thermally driven by the temperature control system. After hybridization, the microarray module is read by the In-Check optical reader (OR) in a few seconds.
In-Check system
The microfluidic integration of the PCR and hybridization modules ensures that system operation does not require highly-trained personnel, and greatly simplifies the typical lab procedures. The In-Check system is extremely userfriendly and allows unskilled operators to perform all steps from sample to result.
Top bench
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Cobas product lines Roche Diagnostics
Clonit StMicroelectronics
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ISyPeM. A solu;on to enable drug-‐ based therapeu;c drug monitoring
• Low-sample volumes • Miniaturized reader • Cartridge that can be stored with stable reagents
• Monitoring and prediction interface for clinicians and practitioners • Local or remote
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• Extension of eHealth medical standards • Data integrity
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ISyPeM: A solu;on to enable drug-‐ based therapeu;c drug monitoring
HOMOGENEOUS AND HETEROGENEOUS ASSAY
• Cartridge that can be stored with stable reagents
In vitro selection of capture molecules for drug targets ISyPeM II -‐ G. Cappi
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In vitro selec;on of monoclonal an;bodies for Tacrolimus Antibody phage display
Heinis Lab
Target: tacrolimus
Sangram Kale
Phage-antibodies were successfully selected for Tacrolimus and expressed ISyPeM II -‐ G. Cappi
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In vitro selec;on of DNA-‐ aptamers for Tobramycin
Guiducci Lab
DNA beacon aptamers Capture Selex
F. M. Spiga Now with Creoptix Sensors
Spiga F. M. et al. ACS Comb. Sci. 2015, featured on the cover of May Issue ISyPeM II -‐ G. Cappi
No drug derivatization High affinity Specific Functional in serum 9
Low-‐volume sample prepara;on on chip Plasma extraction from whole blood Renaud Lab
Yield outperforms any passive on-chip method reported to date
David Forchelet
http://dbs-system.ch
FPIA of Tobramycin performed in glass microcapillaries Squared glass capillary
300 µm
Anisotropy
0.260 0.240
reference system
0.220
capillary
0.200 0.180 0.160 0.140 0
2
4
6
8
10
12
Concentra;on [µg/ml]
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Diana Burghelea J-M Segura
Measurement system: miniaturiza;on of an Polarization established assay technique Fluorescence Immunoassay Competitive assay Highly-sensitive Compatible with small volumes
Martial Geiser team
20 cm
25 cm
*
Requires derivatization of drug molecules ISyPeM II -‐ G. Cappi
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Label-‐free quan;fica;on of drug from serum s amples Heterogeneous assay based on binding kinetics
Guiducci Lab
No drug derivatization No additional reagents
Transmission Surface Plasmon Resonance Change of optical properties of the interface
200 nm
Giulia Cappi
Layers of functionalized gold nanoislands Cappi G. et al. Sensors and Actuators 2013
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Hue r eadout o f s urface p lasmonics Compact read-out system
CMOS image sensor Guiducci Lab
Hue readout of surface plasmonics
Cappi G. et al. Analyt. Chem. 2015
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Measurement of binding kine;cs of DNA aptamer-‐Tobramycin Guiducci Lab
Transmission SPR signal
KD = 200 nM, same as extracted from commercial SPR
Cappi G. et al. Analyt. Chem. 2015
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Tobramycin quan;fica;on from ✔ Small molecules serum ✔ Complex Matrix ✔ Small molecules ✖ Complex Matrix
Chang, A. L. et al., Anal. Chem. 2014, 86, 3273−3278.
✖ Small molecules ✔ Complex Matrix
Jang, H. R., et al. Anal. Chem. 86, 814–819 (2013)
Cappi G. et al. Analyt. Chem. 2015
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Suppor;ng clinical interpreta;on EzeCHiel interface Population-based percentiles
Dosing schedule suggestion
Individual concentration vs time profile Yann Thoma
Div. Clin. Pharmac.
Aline Fuchs ISyPeM II -‐ G. Cappi
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Data management Health data Medical DB POC
Data security Dynamic Health Data Aggregation
Alevtina Dubovitskaya
Data Exchange between EzeCHiel and DB of the Medical “translate” from one representation to another
Institution
RSDB
EzeCHiel
MOLIS, CHUV
Dubovitskaya, A. Privacy Preserving Interoperability for Personalized Medicine, in: Swiss Medical Informa;cs, Swiss Society for Medical Informa;cs, Switzerland, 2014 ISyPeM II -‐ G. Cappi
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Popula;on pharmacokine;cs
Meta-analysis. Implementation and validation
Div. Clin. Pharmac.
Aline Fuchs
Aziz Chaouch
Pharmacokinetic models
Br J Clin Pharmacol. 2014 Nov;78(5):1090-‐101
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Acknowledgements
Guiducci Lab
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