Abstract

Leveraging proteins as therapeutics has revolutionized the way we treat diseases. Recent advances in protein biologics and antibody engineering represent promising new treatment strategies for addressing complex etiologies. Thus far, these advances have been restricted to only accessible extracellular proteins, leaving a significant portion of the proteome undruggable. Efficient intracellular delivery of proteins would reveal uncharted therapeutic avenues and cell surface engineering opportunities. This presentation describes combinatorial strategies towards delivering proteins to the cytosol. By modifying a protein of interest through surface esterification or functionalization with a cell penetrating peptide we have observed robust intracellular delivery. In this presentation, I will first describe the development of a functional assay using a model protein to assess the efficacy and localization of delivery agents. Applying these findings to HaloTag has demonstrated the generality of our approach. I will show that our delivery strategy has enabled stable cell surface engineering. This study highlights that multiple intracellular delivery strategies can be used synergistically to enable cytosolic access with functionally active protein/biologic cargo.

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