Advanced analytical techniques are essential for understanding the complex interactions between therapeutic agents and cellular environments, particularly in the context of precision oncology. This work focuses on the evaluation of the use of two different therapeutic approaches: the use of nanocarriers for cisplatin transport inside the cells [1] and the evaluation of immunotherapy as emerging approaches for cancer management. Immunotherapies represent a broad and rapidly growing group of therapies having a substantial impact on cancer outcomes. Their strength is in their potential to activate the immune system to specifically target cancer cells without the broadly damaging side effects of many conventional chemotherapeutics. Thus, in this presentation, the main objective is to optimize and compare two powerful analytical techniques: sequential single-cell ICP-MS (sc-ICP-MS) using a quadrupole-based mass analyser and simultaneous mass cytometry using a time-of-flight ICP-MS for multiparametric single cell analysis (CyTOF) for the two types of studies. These instrumental approaches will be studied for their ability to provide detailed insights into cellular uptake, distribution, and drug delivery efficacy.

CyTOF is a simultaneous multiparametric single cell analysis technique that enables the simultaneous analysis of multiple cellular markers using mainly lanthanide elements as labels, making it particularly suited for complex biological systems. In this study, the cells are labelled with metal-conjugated antibodies targeting markers such as HER2 and the transferrin receptor TfR1. These markers allow for detailed profiling of cell populations and their nanoparticle interactions. On the other hand, sc-ICP-MS with (triple) quadrupole instruments offers precise quantification of individual elements within cells with extremely high sensitivity. Additionally, triple quadrupole ICP-MS systems offer the possibility to include naturally present elements, such as phosphorus or iron, as interesting cell markers. The main limitation of this instrumentation, however, is given by its sequential measurement approach, which restricts multiplexing capabilities.

This study involves a comprehensive optimization of both techniques, including the preparation and storage of biological samples, calibration of instrument sensitivity for multiple elements, and the assessment of measurement conditions to ensure reproducibility.

References

[1] D. Turiel-Fernández, L. Gutiérrez-Romero, M. Corte-Rodriguez, J. Bettmer, and M. Montes-Bayón, Anal Chim Acta, vol. 1159, May 2021, doi: 10.1016/j.aca.2021.338356.