TY - JOUR

T1 - Fluorescence Masking Based Multifunctional Quantum Dots’ Assay for HSP90α Interactions Detection

AU - Kishore, Anusha

AU - Fan, Lu

AU - Stahl, Frank

AU - Reichel, Thomas

AU - Krüger, Karsten

AU - Zeilinger, Carsten

N1 - Funding Information: The project was funded by the German Federal Institute of Sport Science (Bundesinstitut für Sportwissenschaft, BISP), project ‘Definition of valid and reliable biomarkers including innovative measurement methods for training control in long-distance running’ (grant number 070503/20-21. Funding Information: We would like to acknowledge Institute of Technical Chemistry, Gottfried-Wilhelm-Leibniz University Hannover for providing infrastructural support and Department of Exercise Physiology and Sports Therapy, Institute of Sports Science, JUSTUS-LIEBIG University, Giessen, Germany, for providing materialistic and financial support.

PY - 2023/2/25

Y1 - 2023/2/25

N2 - 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.

AB - 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.

KW - biotin-ATP

KW - biotin-HSP90 antibody

KW - fluorescence masking

KW - HSP90α

KW - quantum dots

KW - Sav-QD

UR - http://www.scopus.com/inward/record.url?scp=85149950401&partnerID=8YFLogxK

U2 - 10.3390/app13052957

DO - 10.3390/app13052957

M3 - Article

AN - SCOPUS:85149950401

VL - 13

JO - Applied Sciences (Switzerland)

JF - Applied Sciences (Switzerland)

SN - 2076-3417

IS - 5

M1 - 2957

ER -