Absolute Quantification of Drug Vector Delivery to the Cytosol.

Marco Lucchino, Anne Billet, Siau-Kun Bai, Estelle Dransart, Justine Hadjerci, Frédéric Schmidt, Christian Wunder, Ludger Johannes

Angewandte Chemie (International ed. in English) 2021 Jun 25

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Macromolecular drugs inefficiently cross membranes to reach their cytosolic targets. They require drug delivery vectors to facilitate their translocation across the plasma membrane or escape from endosomes. Optimization of these vectors has however been hindered by the difficulty to accurately measure cytosolic arrival. We have developed an exceptionally sensitive and robust assay for the relative or absolute quantification of this step. The assay is based on benzylguanine and biotin modifications on a drug delivery vector of interest, which allow, respectively, for selective covalent capture in the cytosol with a SNAP-tag fusion protein and for quantification at picomolar sensitivity. The assay was validated by determining the absolute numbers of cytosolic molecules for two drug delivery vectors: the B-subunit of Shiga toxin and the cell-penetrating peptide TAT. We expect this assay to favor delivery vector optimization and the understanding of the enigmatic translocation process. © 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.

Citation

Marco Lucchino, Anne Billet, Siau-Kun Bai, Estelle Dransart, Justine Hadjerci, Frédéric Schmidt, Christian Wunder, Ludger Johannes. Absolute Quantification of Drug Vector Delivery to the Cytosol. Angewandte Chemie (International ed. in English). 2021 Jun 25;60(27):14824-14830