Zusammenfassung
Immunoassays kamen erstmals in den 1950er Jahren zur Identifizierung und Quantifizierung von Analyten zum Einsatz, wobei der ELISA (engl.: Enzyme-Linked Immunosorbent Assay) erstmals 1971 von Engval und Perlmann beschrieben wurde. Seitdem hat sich das Anwendungsspektrum ständig erweitert, und mittlerweile sind Immunoassays nicht mehr aus der klinischen Forschung und Arzneimittelforschung weg zu denken. Neben dem klassischen kolorimetrischen ELISA setzen sich weitere Nachweistechnologien durch. Diese ermöglichen nicht nur eine höhere Empfindlichkeit, sondern erweitern auch den Anwendungsbereich der Assays. In diesem Kapitel werden einige vielversprechende Mikroplattenreader-basierte Systeme vorgestellt, die alternativ zu klassischen Methoden zur Identifizierung und Quantifizierung von Analyten und zur Untersuchung möglicher Wechselwirkungen eingesetzt werden können.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Literatur
A Practical Guide to working with AlphaScreen. https://www.urmc.rochester.edu/MediaLibraries/URMCMedia/hts/documents/AlphaScreenPracticalGuide.pdf (2004).
Campbell, J. & Carney, O.: Time Resolved Fluorescence (TRF) immunoassay in 384-well format using a matched antibody pair kit and the PHERAstar® FSX. (2020).
Christott, T. et al. AN320: PHERAstar measures AlphaScreen assay to develop selective inhibitors for the human YEATS domains. https://www.bmglabtech.com/pherastar-measures-alphascreen-assay-to-develop-selective-inhibitors-for-the-human-yeats-domains/ (2018).
Coussens, N. P. et al. Compound-Mediated Assay Interferences in Homogenous Proximity Assays. Assay Guidance Manual (Eli Lilly & Company and the National Center for Advancing Translational Sciences, 2004).
Degorce, F. et al. HTRF: A technology tailored for drug discovery – A review of theoretical aspects and recent applications. Current Chemical Genomics vol. 3 22–32 (2009).
Diamandis, E. P. Immunoassays with time-resolved fluorescence spectroscopy: Principles and applications. Clin. Biochem. 21, 139–150 (1988).
Engvall, E. & Perlmann, P. Enzyme-linked immunosorbent assay (ELISA) quantitative assay of immunoglobulin G. Immunochemistry 8, 871–874 (1971).
Förster, T. Energy migration and fluorescence. J. Biomed. Opt. 17, 1 (2012).
Franklin, T. G. & Pruneda, J. N.: Ubiquitination monitoring in real-time: the fluorescence polarization-based method UbiReal. https://www.bmglabtech.com/ubiquitination-monitoring-in-real-time-the-fluorescence-polarization-based-method-ubireal/ (2020).
George, J. & Norey, C.: Use of CyDye fluors for improved FRET protease assays on a BMG LABTECH fluorescence microplate reader. https://www.bmglabtech.com/use-of-cydye-fluors-for-improved-fret-protease-assays-on-a-bmg-labtech-fluorescence-microplate-reader/ (2007).
Homogeneous Assay. in Encyclopedic Reference of Genomics and Proteomics in Molecular Medicine 814–814 (Springer Berlin Heidelberg, 2006). https://doi.org/10.1007/3-540-29623-9_7359.
de Jersey, J. & Martin, R. B. Lanthanide Probes in Biological Systems: The Calcium Binding Site of Pancreatic Elastase As Studied by Terbium Luminescence. Biochemistry 19, 1127–1132 (1980).
Joseph R. Lakowicz. Principles of Fluorescence Spectroscopy. (Springer Science & Buisiness Media, 2013).
Krumm, A., Georgi, A., Schiele, F. & Fernández-Montalván, A.: Binding kinetics: high throughput assay for kinase inhibitors. https://www.bmglabtech.com/de/binding-kinetics-high-throughput-assay-for-kinase-inhibitors/ (2020).
Mattiroli, F., Gu, Y. & Luger, K.: Using FRET-based Measurements of Protein Complexes to Determine Stoichiometry with the Job Plot. https://www.bmglabtech.com/de/using-fret-based-measurements-of-protein-complexes-to-determine-stoichiometry-with-the-job-plot/ (2019).
Morrison, L. E. Time-resolved detection of energy transfer: Theory and application to immunoassays. Anal. Biochem. 174, 101–120 (1988).
Peters, C. & Edwards, B.: Miniaturization of a cell-based TNF-α AlphaLISA assay using Echo liquid handler and the PHERAstar FS. https://www.bmglabtech.com/miniaturization-of-a-cell-based-tnf-a-alphalisa-assay-using-echo-liquid-handler-and-the-pherastar-fs/ (2014).
Riching, K., Landremna, A. & Daniels, D.: Elucidating PROTAC MoA with live cell kinetic monitoring of ternary complex formation and target protein ubiquitination. https://www.bmglabtech.com/de/elucidating-protac-moa-with-live-cell-kinetic-monitoring-of-ternary-complex-formation-and-target-protein-ubiquitination/ (2020).
Soini, E., Lövgren, T. & Reimer, C. B. Time-resolved fluorescence of lanthanide probes and applications in biotechnology. CRC Crit. Rev. Anal. Chem. 18, 105–154 (1987).
Sun, S., Yang, X., Wang, Y. & Shen, X. In vivo analysis of protein–protein interactions with bioluminescence resonance energy transfer (BRET): Progress and prospects. International Journal of Molecular Sciences vol. 17 (2016).
Tsien, R. Y. The Green Fluorescent Protein. Annu. Rev. Biochem. 67, 509–544 (1998).
Ullman, E. F. et al. Luminescent oxygen channeling immunoassay: measurement of particle binding kinetics by chemiluminescence. Proc. Natl. Acad. Sci. U. S. A. 91, 5426–5430 (1994).
Werts, M. H. V. Making sense of lanthanide luminescence. Science progress vol. 88 101–131 (2005).
Xu, Y., Piston, D. W. & Johnson, C. H. A bioluminescence resonance energy transfer (BRET) system: Application to interacting circadian clock proteins. Proc. Natl. Acad. Sci. 96, 151 LP–156 (1999).
Yasgar, A., Jadhav, A., Simeonov, A. & Coussens, N. P. Alphascreen-based assays: Ultra-high-throughput screening for small-molecule inhibitors of challenging enzymes and protein-protein interactions. in Methods in Molecular Biology vol. 1439 77–98 (Humana Press Inc., 2016).
Yi, N. Y. et al.: Monitoring of insulin granule packaging in live cells using homoFRET-FP detection. https://www.bmglabtech.com/monitoring-of-insulin-granule-packaging-in-live-cells-using-homofret-fp-detection/ (2016).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 Springer-Verlag GmbH Deutschland, ein Teil von Springer Nature
About this chapter
Cite this chapter
Pusterla, T., Volz, AC. (2023). Nachweismethoden zur Protein-Interaktion. In: Raem, A.M., Rauch, P. (eds) Immunoassays. Springer Spektrum, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-62671-9_14
Download citation
DOI: https://doi.org/10.1007/978-3-662-62671-9_14
Published:
Publisher Name: Springer Spektrum, Berlin, Heidelberg
Print ISBN: 978-3-662-62670-2
Online ISBN: 978-3-662-62671-9
eBook Packages: Life Science and Basic Disciplines (German Language)