Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses IZI-BB
Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses IZI-BB

Biomarker Validation

Biomarker validation, PopSpot on membrane
© Fraunhofer IZI-BB
PopSpot on membrane

The field of activity includes the development of specific assays for the validation of biomarkers as well as the development and adaptation of assays to different platforms such as microarrays, ELISA, lateral flow systems and beads-based assays in the field of life science, environmental and food analysis. This includes the determination of physicochemical parameters such as kinetic constants (KD) with label-free detection methods (e.g. Biacore, bScreen, Nanotemper), the characterization of interactions e.g. antibody - antigen interactions and the acquisition of parameters that are important for assay development such as the nature or modification of surfaces (e.g. contact angle measurements and ellipsometry). The existing methods and techniques are continuously further developed for (customer-) specific applications. Based on the existing knowledge, specific training and education can be offered to external partners. Methods and techniques used include proteomics techniques such as beads-based methods (Luminex) and microarray but also genomics techniques such as 2nd generation sequencing and various nucleic acid amplification and quantification methods.

 

Applications include system biology projects for validation of potential biomarkers, kinetic analysis and characterization of antibodies, quantification of specific markers in serum samples such as cytokines and miRNAs, and development of point-of-need applications e.g. determination of microbial loads in environmental samples.

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  • Fabrication (spotting) of customized DNA, peptide and protein microarrays.
  • Spotting on various materials, e.g. glass, plastic, membranes, microtiter plates, printed circuit boards, etc.
  • Benchmarking of various contact and non-contact spotters to select the optimal system (reference laboratory for liquid dispensing systems)
  • Execution of customized microarray experiments, evaluation and documentation
  • Optimization of experiments by thermodynamic and kinetic measurements
  • Adaptation of existing assay formats to other methods like microarrays, lateral flow assay but also ELISA
  • Development and establishment of assays for ELISA and microarrays
  • Colorimetric, fluorescence and electrochemical detection systems
  • Serum screening for the identification of disease-associated antibody profiles
  • Detection and validation of (potential) biomarkers in body fluids
  • Epitope mapping of antibodies
  • Bead-based assays (Luminex platform)
  • Sequencing including sample preparation and data analysis and interpretation

  • ELISA
  • Lateral Flow Assay
  • Label-free assays: Biacore, bScreen, Nanotemper
  • Liquid handling and dispensing systems
  • Various microarrays (production, assay development and evaluation)
  • Various detection methods (e.g. optical, fluorescence, calorimetric)

  • Bio-Plex® 3D Suspension Array System
  • Non-contact liquid handling systems such as I2-400
  • bScreen
  • Contact angle measurements, ellipsometry
  • Nanotemper
  • MTP reader

  • Blazquez-Navarro, A., Bauer, C., Wittenbrink, N., Wolk, K., Sabat, R., Dang-Heine, C., Neumann, S., Roch, T., Wehler, P., Blazquez-Navarro, R., Olek, S., Thomusch, O., Seitz, H., Reinke, P., Hugo, C., Sawitzki, B., Babel, N., & Or-Guil, M. (2022). Early prediction of renal graft function: Analysis of a multi-center, multi-level data set. Current Research in Translational Medicine, 70(3), 103334. https://doi.org/10.1016/j.retram.2022.103334
  • Pellegrino, A., Mükusch, S., Seitz, V., Stein, C., Herberg, F. W., & Seitz, H. (2022). Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein. Medical Sciences (Basel, Switzerland), 10(4). https://doi.org/10.3390/medsci10040063
  • Blazquez-Navarro, A., Bauer, C., Wittenbrink, N., Wolk, K., Sabat, R., Dang-Heine, C., Neumann, S., Roch, T., Wehler, P., Blazquez-Navarro, R., Olek, S., Thomusch, O., Seitz, H., Reinke, P., Hugo, C., Sawitzki, B., Babel, N., & Or-Guil, M. (2021). Early prediction of renal graft function: Analysis of a multi-centre, multi-level data set (Vol. 20). https://doi.org/10.1101/2021.01.04.20248473
  • Blazquez-Navarro, A., Dang-Heine, C., Wehler, P., Roch, T., Bauer, C., Neumann, S., Blazquez-Navarro, R., Kurchenko, A., Wolk, K., Sabat, R., Westhoff, T. H., Olek, S., Thomusch, O., Seitz, H., Reinke, P., Hugo, C., Sawitzki, B., Or-Guil, M., & Babel, N. (2021). Risk factors for Epstein-Barr virus reactivation after renal transplantation: Results of a large, multi-centre study. Transplant International : Official Journal of the European Society for Organ Transplantation, 34(9), 1680–1688. https://doi.org/10.1111/tri.13982
  • Morschheuser, L., Mükusch, S., Riedner, M., Seitz, H., & Rohn, S. (2016). High-performance thin-layer chromatography as a fast screening tool for phosphorylated peptides. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 1008, 198–205. https://doi.org/10.1016/j.jchromb.2015.11.055
  • Schumacher, S., & Seitz, H. (2016). A novel immunoassay for quantitative drug abuse screening in serum. Journal of Immunological Methods, 436, 34–40. https://doi.org/10.1016/j.jim.2016.06.004
  • Schumacher, S., & Seitz, H. (2016). Quality control of antibodies for assay development. New Biotechnology, 33(5 Pt A), 544–550. https://doi.org/10.1016/j.nbt.2016.02.001
  • Hahn, M. B., Solomun, T., Wellhausen, R., Hermann, S., Seitz, H., Meyer, S., Kunte, H.-J., Zeman, J., Uhlig, F., Smiatek, J., & Sturm, H. (2015). Influence of the Compatible Solute Ectoine on the Local Water Structure: Implications for the Binding of the Protein G5P to DNA. The Journal of Physical Chemistry. B, 119(49), 15212–15220. https://doi.org/10.1021/acs.jpcb.5b09506
  • Schumacher, S., & Seitz, H. (2015). Multiplex approach for an immunological detection of drug abuse: a validation study. Toxichem. Krimtech, 82, 151-154.
  • Seitz, H., & Schumacher, S. (2015). Biomarker Validation. Wiley. https://doi.org/10.1002/9783527680658
  • Schumacher, S., Muekusch, S., & Seitz, H. (2015). Up-to-Date Applications of Microarrays and Their Way to Commercialization. Microarrays (Basel, Switzerland), 4(2), 196–213. https://doi.org/10.3390/microarrays4020196