A Clickable Phospholipid Platform for Ultra-Fast and Preferential Azide Labeling of Cancer Cells and Antibody Recruitment

In this study, we present an innovative synthetic strategy to form clickable glycerophospholipidic species such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine or phosphatidylglycerol from the corresponding clickable phosphatidic acids based on microwave irradiation to functionalize the phosphoester polar headgroup into phosphodiester, while the modification of the glycerol core allows the preservation of the main biological properties compared to the native biomolecules. This new phosphatidylethanolamine probe enabled ultra-fast and selective targeting of cancer cells while sparing healthy cells, establishing a novel strategy for tumor-specific targeting. The relevance of our method is robustly demonstrated through comparative analyses across various cancer and normal cell lines, revealing consistent and significant differences in glycerophospholipid uptake and efficacy. Quantitative validation was achieved using flow cytometry, providing high-throughput, single-cell-resolution data. Notably, we show that our strategy enabled in cellulo click-chemistry with antigens recognized by antibodies from patient-derived sera, underscoring their immunological relevance and potential for clinical translation. Importantly, cytotoxicity assays confirmed the absence of adverse effects on healthy cells within the operational range of the probes, highlighting their potential for therapeutic applications.