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The peptides were cleaved from the resins using a solution of 95% trifluoroacetic acid, 2.5% H2O, and 2.5% tetraisopropylsilane (TIPS). The Fmoc-protected peptides were purified by HPLC (1260 Infinity, Agilent Technologies) equipped with a C8 column and isolated by lyophilization from the HPLC fractions.
The peptides were conjugated to a polymer to form pH- and redox-responsive coacervate microdroplets. This allowed a wide range of macromolecules to be rapidly recruited into the coacervates, including small peptides, enzymes as large as 430 kDa, and mRNAs, bypassing classical endocytic pathways. The resulting coacervate-mediated delivery bypasses the need for toxic transfection agents, and showed high bioactivity and selectivity. Y.S. designed and performed the experiments, analysed data, and wrote the manuscript. S.Y.L. helped with the EGFP and stapled peptide delivery studies.