Fluorescence Masking Based Multifunctional Quantum Dots’ Assay for HSP90α Interactions Detection

authored by: Anusha Kishore, Lu Fan, Frank Stahl, Thomas Reichel, Karsten Krüger, Carsten Zeilinger
Abstract: Featured Application: The quantum dots-based assay described here uses fluorescence masking to detect protein–ligand and protein–protein interactions on a glass slide in a multifunctional way. This is used here for the stress protein HSP90α in purified form and in cell lysate. It can be further used for other different proteins. HSP90α is one of the most common stress proteins in cells; hence, it is a good target for developing drugs and testing systems for cancer or physical stress levels in humans. Streptavidin conjugated quantum dots (Sav-QDs) are widely used as fluorophores for biosensing to overcome chemical labelling problems. In this work, we have attempted to develop a multifunctional and robust assay for HSP90α. The detection technique was based on the masking of the fluorescence of spotted Sav-QDs on nitrocellulose chips (NC). Biotinylated ligand/antibody attaches to the spotted Sav-QD and then HSP90α is attached, which causes the masking of fluorescence. The masking of fluorescence was used to detect protein–ligand interactions, the effect of inhibitors, protein–protein interactions, and the presence of protein in the biological sample. The load of detection (LoD) of the assay lies in the nano molar range, making it a sensitive assay. The results from the experiments suggest that the used approach is promising for developing a multifunctional, robust, and sensitive assay for proteins that can be used for point-of-care detection in complex biological samples.
Organisation(s): Centre of Biomolecular Drug Research (BMWZ)
Institute of Technical Chemistry
External Organisation(s): Justus Liebig University Giessen
Type: Article
Journal: Applied Sciences (Switzerland)
Volume: 13
ISSN: 2076-3417
Publication date: 25.02.2023
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: General Materials Science, Instrumentation, General Engineering, Process Chemistry and Technology, Computer Science Applications, Fluid Flow and Transfer Processes
Sustainable Development Goals: SDG 3 - Good Health and Well-being
Electronic version(s): https://doi.org/10.3390/app13052957 (Access: Open)