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Protein-containing molecular organized thin film materials with enhanced characteristics for bioanalytical applications and sensors

Objective


Major project's outcome are:
- a piezoelectric immunosensor, which can be used for the detection of small quantities of analytes;
- a prototype phosphorimeter;
- production of electrodes including electrochemical mediators (deposited in an ordered structure) and enzymes;
- production of screen printed electrodes incorporating thermophilic enzymes;
- production of two enzymes from thermophilic bacteria with enhanced stability and with activity superior to mesophilic analogues both at high and room temperatures;
- production of a new NADH-oxidising compounds with amphiphilic properties;
- stabilisation of enzyme structure and activity with the use of polyalcohol molecules, and development of new methodology for analysis of mechanism of stabilisation;
- an innovative spectroscopic technique based on protein phosphorescence to determine protein structure;
CONFIDENTIAL
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Objectives and content
The project has the objectives:
to produce novel functionally active materials based on
thin films of biological molecules, with enhanced
properties, to be used in applications where principles
of bioanalysis are utilised;
to develop technologies, including Langmuir-Blodgett
related techniques to reproducibly yield such materials;
to demonstrate the improved functionality of new
materials for specific chosen industrially relevant
applications within sensing elements of chemical and biosensors, for various bioanalytical operations such as
clinical analysis, drug screening, monitoring, as well as
biotransformation processes.
The project handles the priority research tasks of the
sub-area 2.1 Materials Engineering: 2.1.4.M, 2.1.2.L and
2.1.3.L.
The innovation of the present project consists in three
main points. The first one is based on the application
of a recently proposed method of LB film deposition which
enables easy inclusion of protein layers in the desired
position of the layered structure to be deposited without
any contact of proteins with air medium, thus preventing
protein denaturation and protecting protein layers by
monolayer coatings afterwards.
The second innovation consists in the combination of the
developed methods of stabilisation of protein functional
properties utilising polyelectrolytes and/or
polyalcohols, with the above technique of film
deposition. This approach will enable to introduce
particular stabilising agents into the film structure at
every and any step of monolayer deposition and protein
adsorption.
The third innovative approach is the application of
protein refolding technique to increase the activity of
proteins both in the initially created film structure and
after ageing of the latter. The refolding approach is
based on a novel protein with ATP-dependent activity and
the disulphide bond-forming activity.
The combination of the above approaches permits also to
control the orientation of protein molecules in the film,
their reciprocal orientation with respect to lipid and
stabiliser molecules, in order to control permeability
towards substrates and products as well as accessibility
of active sites.
The expected increase in storage stability and shelflife
might reach 300 %, in sensitivity - 30 %, in stability
versus solvents - at least 50 %, in the possibility of
multiple usage - by 50%. Moreover, the activity of
enzymes immobilised in conventional carrier might be
increased by 50 % by utilising the refolding technique,
where applicable.
The industrial objectives will be warranted by two SMES
taking part in the project in accordance with the
business strategy of these companies.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

Consorzio Tecnobiochip Sarl
Address
Via Della Marina
57030 Marciana Marina
Italy

Participants (4)

Applied Enzyme Technology Limited
United Kingdom
Address
Woodhouse Lane
LS2 3AR Leeds
NATIONAL RESEARCH COUNCIL OF ITALY
Italy
Address
Area Della Ricerca, Via Alfieri 1
56010 Mezzana
UNIVERSITAT ROVIRA I VIRGILI
Spain
Address
Avinguda Dels Paisos Catalans 26
43007 Tarragona
WAGENINGEN UNIVERSITY
Netherlands
Address
4,Hesselink Van Suchtelenweg 4
6703 CT Wageningen