A fluorescence based turn OFF – ON detection of cancer

Authors: Dominik Michálek 1    Zuzana Garaiová 2    Sylwia Michlewska 3    Maksim Ionov 3    Maria Bryszewska 3    Natalia Sanz del Olmo 4    Francisco Javier de la Mata 4    Martin Kopáni 5    Iveta Waczulikova 2    Tibor Hianik 2   
1 Faculty of Medicine Comenius University, Bratislava, Slovakia    2 Faculty of Mathematics, Physics and Informatics Comenius University, Department of Nuclear Physics and Biophysics, Bratislava, Slovakia    3 Faculty of Biology and Environmental protection, University of Lodz, Department of General Biophysics, Laboratory of Microscopic Imaging and Specialized Biological Techniques, Lodz, Poland    4 Department of Organic and Inorganic Chemistry, and Research Institute in Chemistry” Andrés M.Del Río” (IQAR), University of Alcalá, Madrid, Spain    5 Faculty of Medicine Comenius University, Institute of Medical Physics, Biophysics, Informatics and Telemedicine, Bratislava, Slovakia   
Year: 2020
Section: Biophysics, mathematical modeling, biostatistics
Abstract No.: 2046
ISBN: 978-80-972360-6-9

High-quality and accurate biodetection methods ensuring effective disease diagnostic represents still a major challenge. Recently, a new detection mode based on “turn OFF – ON” fluorescence change (quenching-recovery) through a Fluorescence resonance energy transfer (FRET) has attracted a considerable research interest [1]. FRET is a distance-dependent radiation-lees transfer of energy from an excited molecular donor fluorophore to an acceptor molecule, which can be used to investigate molecular level interactions [2].

In this study, turn OFF – ON fluorescence method based on surface energy transfer mechanism for detection of leukemic Jurkat T-cells was investigated. We worked with fluorescently labeled carbosilane dendrimers based on ruthenium FITC-CRD13 (acts as donor) possessing anticancer properties [3] and graphene oxide GO (acts as acceptor) that is known as an effective fluorescence quencher [4].  Firstly, complexes of FITC-CRD13 + GO were prepared, and fluorescence intensity measured. Due to the proximity (adsorption) of dendrimer to GO, fluorescence intensity was quenched (fluorescence OFF). The fluorescence intensity of the FITC-CRD13 dendrimer (100 nM) reached the quenching maximum by the concentration of 30 μg/ml GO. Subsequently, the recovery of the fluorescent signal (fluorescence ON) upon the interaction of FITC-CRD13 + GO composite with Jurkat T-cells was examined. Upon the interactions with 106 cells/ml, the fluorescence intensity increased approximately 6-times in comparison with control sample (no cells). An increase in fluorescence was also reported for concentration 105 cells/ml, reaching an approximately 4-fold increase in signal. We suppose that much stronger affinity of dendrimer to cellular receptors takes place and leads to the desorption of dendrimer from the GO structure, which allows the regaining of the FITC-CRD13 fluorescence. To verify specificity of the studied platform, interaction with other cell lines needs to be investigated, too. Obtained results indicate the potential of turn OFF – ON fluorescence platform in the field of biodetection and targeted therapy of cancer.

This work was supported by the Slovak Research and Development Agency, APVV (Projects No. SK-PL-18-0080, APVV-14-0267, SK-BY-RD-19-0019), COST CA17140, VEGA 1/0756/20 and by Polish Ministry of Science and Higher Education together with joint project EUROPARTNER: Strengthening and spreading international partnership activities of the Faculty of Biology and Environmental Protection for interdisciplinary research and innovation of the University of Lodz, which is financed by NAWA International Academic Partnership Programme.
[1] Tan J, Lai Z, Zhong L, Zhang Z, Zheng R, Su J, Huang Y, Huang P, Song H, Yang N, Zhou S, Zhao Y. Nanoscale Res Lett.1;13(1):66, (2018)
[2] S. Zadran, S. Standley, K. Wong, E. Otiniano, A. Amighi, M. Baundry, Applied Microbiology and Biotechnology 96, 895 -902, (2012)
[3] S. Michlewska, M. Kubczak, M. Maroto-Díaz, N.   Sanz Del Olmo, P. Ortega, D Shcharbin, R. Gomez Ramirez, F. Javier de la Mata, M. Ionov, M. Bryszewska, Biomolecules 9(9), (2019)
[4] E. Morales‐Narváez, A. Merkoçi, Advanced Materials 24(25), 3298-3308 (2012)