MICROSENSOR: Construction of miniaturised free flow electrophoresis (mffe) systems incorporating dedicated sensors for real-time analysis of food contaminants

• Forschungsstelle: EU FRP
• Projektnummer: 99.0065
• Projekttitel Englisch: MICROSENSOR: Construction of miniaturised free flow electrophoresis (mffe) systems incorporating dedicated sensors for real-time analysis of food contaminants

Schlüsselwörter (Englisch)

Free flow electrophoresis; biosensors; grating couplers; integrated optics

Alternative Projektnummern (Englisch)

EU project number: QLRT-1999-00343

Forschungsprogramme (Englisch)

EU-programme: 5. Frame Research Programme - 1.1.1 Food, nutrition and health

Kurzbeschreibung (Englisch)

See abstract

Partner und Internationale Organisationen (Englisch)

Coordinator: LFRA, Leatherhead (UK)

Abstract (Englisch)

Objectives: It is the strategic objective of this project to test the practical feasibility of miniaturised free flow electrophoresis (mFFE) systems with downstream dedicated sensors for the real-time measurement of three selected analytes (e.g. mycotoxin, markers for recombinant bovine somatotrophin, rBST, and Listeria) as a tool to address the problems of food safety. The main intermediate objectives are:· To design and engineer, to pre-determined specifications, analytical miniaturised FFE systems on suitable substrates (e.g. silicon and glass).· To investigate and optimise the performance of generic mFFE systems for the selected analytes initially from simple aqueous preparations and then complex food matrices.· To determine the isolated analytes downstream using modern optical biosensor technology in comparison with current accepted methods.· To examine the potential of mFFE systems for direct measurement of the analytes (e.g. line sample measurement for process monitoring and control in Quality Assurance).· To critically consider exploitation and dissemination plans of the envisaged technology resulting from this research project. The specific objectives set for this reporting period were:· Continue the development of a pre-prototype microfabricated FFE chip device (e.g. volume £100 ml) · Design and performance test pre-prototype mini-fabricated FFE device and technology transfer to the food laboratory· Continue the development of an integrated software package and flow cell module for ASI's biosensor and technology transfer to the food laboratory· When available, set-up the miniaturised FFE and sensor instrumentation in the food laboratory for the development of practical protocols Results: The literature reviews on the applications of FFE and biosensors in the agrifood industry have been published as LFRA reports. Further external publication of the biosensor paper has been secured in the Trends in Analytical Chemistry (TRAC) Journal. Work has continued on the development of miniaturised FFE systems. Standard procedures for microfabrication of polydimethylsiloxane (PDMS) chip devices have been established, permitting the rapid and cost-effective production of multiple copies. The first-design prototype of a microchip, with media flow regulated by a vacuum pump, has been fabricated for use with a linear array UV detector. The integrity and functionality of the structure has been successfully demonstrated by resolving the fluorescent compounds fluorescein and rhodamine. The microchip magnet design concept has been patented. Work has also continued on the development of prototype mini-FFE chambers for the separation of Listeria and IGF-1. Continuous isotachophoresis was used to resolve Listeria and Micrococcus from a mixed suspension. Sample capacity was increased using simultaneous manifold separations. The robustness of the mini-chambers was improved by developing a new supporting frame to equalise the pressure across the chambers. Following successful reliability trials, the mini-FFE instrument was technology transferred to the food laboratory for the development of practical protocols. Free flow zone electrophoresis and isotachophoresis have since been used to determine the characteristic profiles of Listeria, Micrococcus and E.coli using the mini-FFE instrument. Work has continued on the development of suitable detection systems for use with the mFFE. Work was instigated on development of SPR sensor based assay procedures for Listeria. Selective binding profiles of antibody and bacterial cells have been demonstrated using gold coated glass slides of a commercial optical biosensor. The ASI biosensor has been tested for stability, and recently transferred to the food laboratory for protocol development. Milestones: · Brief review of scientific and technological basis of FFE and sensor techniques (month 6) ACHIEVED · Establish first design microfabricated FFE system (month 8) ACHIEVED· Establish first design minifabricated FFE system (month 9) ACHIEVED· Advance software package for the optical biosensor system (month 15) ACHIEVED· Practical protocol for an analyte based on mFFE and the associated sensor (month 26) IN PROGRESS Benefits and Beneficiaries: The project aims to deliver a range of dedicated novel instrumentation for the analysis of selected food contaminants. It is envisage that the systems(s) will be generic and modular to allow for a wide variety of applications to be developed. In addition to the target analytes selected in this project, the systems will have scope in analysis and characterisation of biomolecules and particles in many other industries (e.g. Medicare, Water and Agriculture). The modular nature of the systems is expected to form the basis of automated integrated analytical instrumentation. Future Actions: · Continue construction of new micro- and mini-FFE modules, and performance test pre-prototype instruments using selected analytes. · Continue development and integration of advanced software into an automated biosensor system.· Continue development of practical methods for target analytes in food systems.· Continue dissemination of information, as and where appropriate.

Datenbankreferenzen (Englisch)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 99.0065