WP2: Design and modulation of new biomaterials as sensor element.

Objectives:

• Proteins and small molecules preparation. Structure-function characterization using available experimental data.

• High- throughput screening of potential agonist, antagonist and inhibitor molecules.

• In silico protein engineering.

Expected results:

• Determine molecular parameters that correlated with properties to be improved.

• Best small molecules selected to be tested experimentally.

• Set of TRP, MscL/MscS, ASIC proteins to be tested experimentally

Supervisors: Laura Masgrau, Joan Torras

Secondments:

• University of Teramo (Italy). FRET measurements of membrane binding affinity.

• Polytechnic University of Catalonia (Spain), QM/MM-MD study of some agonist/antagonist in ion channels.

Doctoral programme: Polymers and Biopolymers at Polytechnic University of Catalonia.

WP2: Design and modulation of new biomaterials as sensor element.

Objective:

• Selection and modification of thermosensitive ion channels to be used as thermosensors.

Expected results:

• A thermosensitive ion channel which only response to Temperature and is insensitive to other stimuli’s

Supervisor: Thomas Voets

Secondments:

• University of Teramo (Italy). Live cell imaging of TRPV1 ion channels.

• Zymvol Biomodeling S.L. (Spain). In silico study of TRP mutagenesis.

Doctoral programme: Cognitive and Molecular Neurosciences at KU Leuven.

WP2: Design and modulation of new biomaterials as sensor element.

Objectives:

• Identifying and characterization of ASIC mutant with sustained current

• Protein expression and purification of the selected mutants.

• Measuring electrical properties measurements on planar lipid bilayers with ion channels.

Expected results:

• Full characterization of several mutants with sustained current and wide pH range.

• Expression of functional ion channels with high yield and purity.

• Integration of the acid-sensing ion channels in different polymers to evaluate the most effective polymer to be used as sensor skeleton.

Supervisor: Stephan Kellenberger, Ivan Gautschi

Secondments:

• École Polytechnique Fédérale de Lausanne (Switzerland). ASIC Protein expression and purification.

• University of Warsaw (Poland). Electrical properties measurements on planar lipid bilayers with ion channels.

• Zymvol Biomodeling S.L. (Barcelona). In silico mutagenesis study of ASIC proteins.

Doctoral programme: Doctoral School of Life Sciences at University of Lausanne.

WP2: Design and modulation of new biomaterials as sensor element.

Objectives:

• Protein stabilization into the lipid bilayer and development of specific synthetic membranes. 


• Expression and purification of selected membrane proteins.

Expected results:

• More efficient binding of the selected protein on the developed nanosensors.

• Ex vivo isolation from different cells of specific membranes for stabilization of the purified proteins. 


• Production and use of different unilamellar vesicles for membrane binding affinity studies.


• Integration of the cholesterol sensing polypeptide with different supports (lipid bilayer and/or conducting polymer) to study the best options using also different systems to increase the signal

Supervisor: Enrico Dainese, Sergio Oddi

Secondments:

• University of Groningen (Netherlands). Membrane reconstitution and microscopy.


• University of Lausanne (Switzerland). Measurement of ASIC modulation by small compounds. 


• 4Dcell SAS (France). Training of Organ on Chip.

Doctoral programme: University of Teramo.

WP2: Design and modulation of new biomaterials as sensor element.

WP3: Design and optimization of a biodegradable sensor skeleton and electrodes.

Objectives:

• Generation of hybrid vesicular systems based on amphiphilic block copolymers and lipids.

• Reconstitution of simple peptides (e.g. pore-forming antimicrobial peptides) and membrane proteins (e.g. MscL, wild type and mutants) in hybrid vesicles and polymersomes.

• Determine the lipid dependence of MscL and then compare their performance with that of hybrid membrane vesicles.

Expected results:

• Stable and functional proteo- hybrid vesicles suitable for biosensing.

• New insights in the gating mechanism of MscL (wt and mutants).

Supervisor: Bert Poolman, Giovanni Maglia

Secondments:

• VIB-KU Leuven (Belgium). Electrophysiology analysis of ion channels.

• Zymvol Biomodeling S.L. (Spain). In silico study of MscL/MscS mutagenesis.

• University of Warsaw (Poland). Nanoscale visualization of ion channels in lipid bilayer.

Doctoral programme: University of Groningen.

WP3: Design and optimization of a biodegradable sensor skeleton and electrodes.

Objectives:

• Fabricate functionalized and perforated planar polymer membranes as a mechanical support to tether lipid bilayers.

• Prepare functionalized micro- and nanospheres of polymers to stabilize the lipid vesicular system.

• Identify the relationship between polymer structure and lipid membrane composition to get more system stability.

Expected results:

• Effective anchoring of lipid bilayers over polymer support without compromising the functions of the implantable sensor.

• Deposition of lipid layer on the surface of functionalized microsphere for biosensing applications.

Supervisor: Joan Torras, Elaine Armelin

Secondments:

• University of Warsaw (Poland). Stable planar lipid bilayer building.

• University of Groningen (Netherlands). Preparation of vesicles with PLA skeleton.

• 4DCell SAS (France). Biodegradability study in novo testing platform.

Doctoral programme: Polytechnic University of Catalonia.

WP3: Design and optimization of a biodegradable sensor skeleton and electrodes.

WP4: Sensor integration and testing.

Objectives:

• Design and synthetize new biodegradable conducting polymers optimizing their electrical conductivity and biodegradability.

• Obtaining novel polymeric conductive materials with high biocompatibility and conductivity, and low toxicity.

Expected results:

• Development of flexible electrodes based on biodegradable conducting polymers and/or bioresorbable metals.

• Integration of a sensor skeleton made of a nanoperforated nanomembrane and flexible electrodes.

Supervisor: Joan Torras, Francesc Estrany

Secondments:

• MECWINS (Spain). Building electronic circuits for sensor signal treatment.

• 4DCell SAS(France). Signal processing technology.

Doctoral programme: Polytechnic University of Catalonia.

WP3: Design and optimization of a biodegradable sensor skeleton and electrodes.

WP4: Sensor integration and testing.

Objectives:

• The project assumes the use of the artificial lipid bilayers as platforms that can provide a suitable environment for the insertion of transmembrane proteins while retaining their native function.

• The project also assumes the characterization of the functionality of the ion channels upon insertion into the lipid bilayers. The response of the channels upon application of external stimuli (change of the pH, temperature, electric field) will be monitored in situ with electrochemical methods combined with surface-enhanced infrared absorption spectroscopy (SEIRAS).

Expected results:

• Develop planar lipid bilayers with fully functional ion channels which activity can be triggered by means of physical or chemical stimuli.

Supervisor: Slawomir Sęk

Secondments:

• University of Lausanne (Switzerland). Training in electrophysiological methods of analysis of ion channels.

• Polytechnic University of Catalonia (Spain). Preparation of PLA membranes for lipid bilayer immobilization.

• 4Dcell SAS (France). Device preparation of cell microenvironment control.

Doctoral programme: University of Warsaw.

WP4: Sensor integration and testing.

Objectives:

• Develop a microfluidic platform that will mimic in vivo conditions to test the behavior of the sensor.

Expected results:

• Microfluidic testing platform with integrated sensors closely resembling the in vivo cellular microenvironment of sensory cells.

Supervisor: Patricia Davidson, Thomas Voets

Secondments:

• VIB-KU Leuven (Belgium). Thermodynamic analysis of temperature sensitivity of on channels.

• University of Warsaw (Poland). Nanomechanical analysis of planar lipid bilayers on sensors.

Doctoral programme: KU Leuven.

WP4: Sensor integration and testing.

Objectives:

• Application of the cholesterol peptide for in vitro cell imaging and collaboration on sensor integration.

Expected results:

• Evaluation of different supports for the cholesterol binding peptide including conducting polymers and nanomechanical sensors.

• Development of cell-live imaging techniques for visualizing the localization, organization, and dynamics of intracellular cholesterol.

• Collaboration for the validation of biosensors as novels tools measuring cholesterol & acidity in living cells.

Supervisor: Enrico Dainese, Sergio Oddi

Secondments:

• Polytechnic University of Catalonia (Spain). Immobilization of a cholesterol polypeptide sensor in conducting polymer.

• Mecwins S.L. (Spain). Cholesterol determination using SCALA technology.

Doctoral programme: University of Teramo.

WP4: Sensor integration and testing.

Objectives:

• Design, development and testing of the associated electronics and signal treatment of the ionic channel sensor, later to be tested in living models.

Expected results:

• Measurement of the ionic channel conductances and action potentials coming from the micropatterned cell samples (muscle and cardiomyocytes).

• Amplification and signal treatment of the incoming signals by integrated electronics.

Supervisor: Patricia Davidson, Olivier Français

Secondments:

• Universite Gustave Eiffel (ESIEE, Paris). Instrumentation and alectronics for biosensors.

• AO Foundation (Switzerland). Animal testing of developed biosensors.

Doctoral programme: Universite Gustave Eiffel.