A Photoactive Supramolecular Complex Targeting PD-L1 Reveals a Weak Correlation between Photoactivation Efficiency and Receptor Expression Levels in Non-Small-Cell Lung Cancer Tumor Models
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Purification of Atezolizumab and Labeling with EITC or AlexaFluor647
2.3. Spectral Properties of the Conjugate and 1O2 Production
2.4. Cell Culture
2.5. Evaluation of PD-L1 Expression
2.5.1. Flow Cytometry
2.5.2. Microscope Setup
2.5.3. Sample Preparation and Acquisition for dSTORM Imaging
2.5.4. dSTORM Image Analysis
2.6. Photodynamic Inactivation of Cultured Cells
2.6.1. Analysis of Cell Viability and Cell Death
2.6.2. ROS Detection
2.6.3. Statistical Analyses
3. Results and Discussion
3.1. Quantitative Determination of PD-L1 Expression Levels
3.1.1. PD-L1 Receptor Density via Flow Cytometry
3.1.2. PD-L1 Receptor Density via dSTORM
3.2. EITC–Atezolizumab Complex for Photo-Immunotherapy
3.2.1. Spectral Properties of EITC–Atezolizumab
3.2.2. Singlet Oxygen Production by EITC–Atezolizumab
3.2.3. Binding Capability of the Conjugate to Cell Lines Expressing PD-L1
3.3. Photoinactivation Treatment with the Conjugate
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Delcanale, P.; Alampi, M.M.; Mussini, A.; Fumarola, C.; Galetti, M.; Petronini, P.G.; Viappiani, C.; Bruno, S.; Abbruzzetti, S. A Photoactive Supramolecular Complex Targeting PD-L1 Reveals a Weak Correlation between Photoactivation Efficiency and Receptor Expression Levels in Non-Small-Cell Lung Cancer Tumor Models. Pharmaceutics 2023, 15, 2776. https://doi.org/10.3390/pharmaceutics15122776
Delcanale P, Alampi MM, Mussini A, Fumarola C, Galetti M, Petronini PG, Viappiani C, Bruno S, Abbruzzetti S. A Photoactive Supramolecular Complex Targeting PD-L1 Reveals a Weak Correlation between Photoactivation Efficiency and Receptor Expression Levels in Non-Small-Cell Lung Cancer Tumor Models. Pharmaceutics. 2023; 15(12):2776. https://doi.org/10.3390/pharmaceutics15122776
Chicago/Turabian StyleDelcanale, Pietro, Manuela Maria Alampi, Andrea Mussini, Claudia Fumarola, Maricla Galetti, Pier Giorgio Petronini, Cristiano Viappiani, Stefano Bruno, and Stefania Abbruzzetti. 2023. "A Photoactive Supramolecular Complex Targeting PD-L1 Reveals a Weak Correlation between Photoactivation Efficiency and Receptor Expression Levels in Non-Small-Cell Lung Cancer Tumor Models" Pharmaceutics 15, no. 12: 2776. https://doi.org/10.3390/pharmaceutics15122776