Nanotechnology based assays for validating protein biomarkers

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The part of the project conducted at Fraunhofer IZI involves selecting immunological parameters in 10 animals, validating these immunological parameters in 100 animals, and developing and validating laboratory tests for immunological parameters for bulk investigation and field validation of the tests on 3 selected farms. PMID: 24870377 Identification and evaluation of novel synovial tissue biomarkers in rheumatoid arthritis by laser scanning cytometry. In the meantime, a cellular biomarker (CD25 in neutrophilic granulocytes) and two protein biomarkers have been identified and validated. [Epub ahead of print] Meinlschmidt P, Brode V, Sevenich R, Ueberham E, Schweiggert-Weisz U, Lehmann J, Rauh D, Knorr D, Eisner P. Fueldner C, Mittag A, Knauer J, Biskop M, Hepp P, Scholz R, Wagner U, Sack U, Emmrich F, Tárnok A, Lehmann J. ELISA, western blot, and peptide or bead arrays (Luminex) are utilized for validation. Stefan Kalkhof also holds the research professorship for the master degree program Bioanalytics at the Coburg University for applied science and arts.High-affinity monoclonal antibodies, which are usually developed in the group, are key tools for these immunochemical assays. This synergy enables the identification and characterization of relevant biomarkers by mass spectrometry, which are then validated in the Department of Therapy Validation using antibody-based techniques and prepared for clinical application.

The project goal is to design a diagnostics platform the enables rapid as well as selective diagnosis of the target protein morphologies from clinical samples.Biomarker discovery is usually done by non-targeted “shotgun” proteomics, using relative quantitation methods to determine protein expression changes that correlate with disease (output given as “up-or-down regulation” or “fold-increases”).MS-based techniques can also perform “absolute” quantitation which is required for clinical applications (output given as protein concentrations).Moreover, the unit aims at the development of single and multiplex assays for their detection.Multi-omics strategies (especially LC-MS based proteomics) are applied for identification.

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