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The BioResponse association is developing a wealth of innovative technical solutions that combine to form the core technology of BioResponse for multiparametric diagnostics, consisting of:
        - Antibody bead assay
        - Cell assay
        - the VideoScan measuring system
        - AKLIDES® as the first commercially available system
        - Real time PCR
        - Microparticles
        - Analysis of bacterial infection in cell culture model
        - Enzyme assays
The multiple uses of the BioResponse technology in diagnostics, bio-analytics and pharma screening is based on the different fields of application like, e.g. protein analytics (antigens, antibodies), DNA analytics or cell analytics with an abundance of applications. This wealth demands the initial focussing of the research association at the most important and most likely successfully applied implementations (infection serology, auto immune diagnostics, gene tests and molecular diagnostics for infectious diseases).

Bead Assay
The bead assay (BA) is one of the forms of the BioResponse technology and allows analysis of proteins, nucleic acids or other molecules. The BA is able to measure different parameters within one reaction vessel, e.g., one well of a microtestplate. Microtestplates consist of 96 reaction vessels, belong to the routine equipment in a diagnostics laboratory and allow for an automatic and parallel processing of patient samples.
    The binding of specific antibodies from the sample to one of the different, immobilized biomolecules can be detected with a fluorescence-labeled detection antibody. The biomolecules in question are bound to fluorescence-encoded microparticles, so that identifying the reacting molecules is possible.

Beadarray with coded particles
The amount of work and materials needed compared to existing immunoassays can be reduced by the factor 10. Up to 20 parameters can be examined at the same time, this is an innovation in a routine laboratory.
    Compared to conventional biochips with xy-coordinates, whose parameters are generally printed in nanoliter amounts on carrier-materials, the so-called 'nanoliter liquid handling' is not applicable. Especially for proteins, this technology is not being mastered to produce repeatable results. The required quality for screening or confirmatory tests in multiparameter diagnostics can only be reached by using the BioResponse technology.
    This promising technology was being developed, tested and brought to a pre-marketable prototype by R&D-grants from the Bundesministerium für Bildung und Forschung (BMBF) from 2004 to 2007 during the 'Wachstumskern BioResponse' phase.
    Through increased sensitivity and specifity on one hand, and a drastic reduction of production costs on the other hand, the Bead-assay is revolutionary in comparison to other test systems. Recently, first products have been introduced into the market.

Reaction scheme of an antibody test for detecting anti-nuclear antibodies (ANA) with the bead assay. Example of an anti-dsDNA-positive sample.

Cell assay
Autoimmune disease is the generic term for diseases whose cause is an out-of-control reaction of the immune system against the body's own tissues. These diseases are known as rheumatic inflammations and are characterized by occurrences of autoantibodies (AAB) that are used as markers for different kinds of disease patterns and can also be used as markers of activity. The qualitative and semi-quantitative detection of AAB in human sera is normally carried out with indirect immunofluorescence (IFT) on human epithelial cells (HEp2-strain) that are being used as a natural antigenic substrate. IFT-testing on HEp2 cells is a sensitive and proven screening assay for determining AAB, and also allows a proposition of certain underlying antigens and associated diseases by recognizing patterns of fluorescence. The conventional visual analysis of the immunofluorescence patterns is disadvantageous because of a high variability of the expertise of the analysing person and the degree of standardization of the lab workflow.
    The measuring system 'Aklides' has been developed, consisting of a robust detection system, including new solutions in hard- and software-based analysis of cells and tissues in combination with a system design based on existing and yet-to-be-developed algorithms to automatically characterize HEp2-immuno-fluorescence.

Cell assay for detection of auto-antibodies on HEp-2 cells

Measuring system Videoscan
The measuring system Videoscan was being developed to analyse microparticles (MP) and cells. The system excels at highly selective, imaging multifluorescence detection.
In order to obtain a number of different colours, respectively colour-intensity combinations for the BA, a very high differentiation of colours is being used, that no other measuring system for parallel detections at present is being able to reach.
Besides being used as a detection systems for analysing MP, the Videoscan technology is suitable for in-vivo or in-situ characterizations of cells and tissues.
The technological prerequisites for the detection and automatic interpretation of cellular patterns of immunostained cells were being established in the project Cell-Chip, using the example of HEp2 cells.
The LoopTag-probe system, patented by the partners, allowed the development of leading pathogen detection kits for microbiological diagnostics of infectious diseases. To date, these tests allow the differentiation of up to three nucleic acid targets in patient samples, the detection of an internal standard as well as the DNA fragments of two different causative agents by melting curve analysis. In doing so, a sensitivity of up to one pathogenic agent genome equivalent per reaction is being achieved.
The microparticles being developed by the PolyAn GmbH consist of polymethylmethacrylate (PMMA). For coupling biomolecules, they can be equipped with a reactive 3D-surface. For the use in flowcytometrics, imaging systems for multiparametric diagnostics like, e.g., the BioResponse-system as well as screening purposes, PolyAn has developed polymeric microparticles consisting of PMMA, which are encoded by two fluorescent dyes.
For every given size there exist up to 20 different bead populations, which can distinguished by the ratio of both integrated dyes.
The integration of several fluorophores and the precise production in classes of size between 2 - 25 µm with a standard deviation in size of less than 2 - 5 % enables the production of a great number of well distinguished bead populations for multiplex-uses. Due to the 3D-functional layer on the particle surface, there is only slight amount of aggregation of beads and a high signal-noise-ratio.

The Videoscan process as fluorescence-spectroscopic detection system for microparticles and cells

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