CELL-BASED BIOSENSORS IN THE 21ST CENTURY: BACK TO THE BASICS OR A QUANTUM LEAP FOR BIOTECHNOLOGY?

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• Prof. Spiridon KINTZIOS, Faculty of Agricultural Biotechnology, Agricultular University of Athens, Greece and EMBIO Diagnostics, Nicosia, Cyprus
• Thursday 07 August 2008, 14:30-15:30
• Mott Seminar Room, Cavendish Laboratory.

If you have a question about this talk, please contact Dr Thanos Mitrelias.

In recent years there has been a rapid increase in the number of diagnostic applications based on biosensors, including live, intact cells, tissues, organs or whole organisms. Cell-based biosensors represent the next revolution in medical diagnostics, offering a number of significant advantages, such as high speed, portability and low cost. The present review focuses on the most successful technologies used for the detection of ultra-low concentrations of bioactive analytes (such as metabolic markers and pathogens) in clinical and other samples. In similar fashion to DNA and protein microarrays, which deliver multiplex detection via the high-density spatial arrangement of molecular recognition elements, arrays of cells at high-density can form the basis of cell-based sensors with extremely high-throughput capability. The expression of receptors of interest within these arrays could yield cell-based sensors with defined specificities. In addition, transfected cell microarrays (such as the CANARY system) composed of high-density arrays of mammalian cells expressing de-fined genes, could be the basis for future high-throughput cell-based protein sensing platforms. The artificial insertion of receptor-like molecules in the cell membrane is an attractive alternative to cell transformation with genes expressing membrane-bound antibodies. This generic approach is called Molecular Identification through Membrane Engineering (MIME). Interaction of MIME cells with target analytes can trigger changes to the cell membrane potential that are measured by appropriate microelectrodes, according to the principle of the Bioelectric Recognition Assay (BERA). BERA is a biosensory method based on a unique combination of a group of cells, whose immobilization in the matrix preserves their physiological functions and measures the expression of the cell interaction with target molecules, through the change in electrical properties. In this way, when a positive sample is added to the probe, a characteristic, ‘signature-like’ change in electrical potential occurs upon contact between the target molecule and the gel matrix. BERA has been used for the detection of a plethora of analytes in humans, animals, plants and agricultural/food commodities in a remarkably specific, rapid (1-2 minutes), reproducible and cost-efficient fashion. The sensitivity of the virus detection with BERA is equal or even better than with advanced immunological, cytological and molecular techniques, such as the reverse transcription polymerase chain reaction (RT-PCR). The BERA biosensor diagnostic system is currently available as a desktop, laboratory-scale prototype that can be operated by both expert and lay users. The commercialization process of the device includes engineering for a more compact, stand-alone unit. The system comprises a consumable miniature biosensor (with integrated circuitry, an immobilization matrix and virus-specifically responding cells), a data acquisition system and a PC (desktop or laptop). One of the major advantages of BERA is the extended storability of the disposable sensors, which is also documented by other research groups. So far, more than 100,000 sensors have been used for screening worldwide. Further advances in microfluidic cell circuits (like HuREL®) and the development of generic methods for the specific detection of target analytes, like CANARY and BERA , along with an increasing investment in automated equipment and user-friendly software, pave the way for the final destination of cell biosensors: the diagnostic laboratory.

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