Synergistic Interplay of Covalent and Non‐Covalent Interactions in Reactive Polymer Nanoassembly Facilitates Intracellular Delivery of Antibodies

Abstract

The primary impediments in developing large antibodies as drugs against intracellular targets involve their low transfection efficiency and suitable reversible encapsulation strategies for intracellular delivery with retention of biological activity.T oa ddress this,w eo utline an electrostatics-enhanced covalent self-assembly strategy to generate polymer-protein/antibody nanoassemblies.Through structureactivity studies,w ed own-select the best performing selfimmolative pentafluorophenyl containing activated carbonate polymer for bioconjugation. With the help of an electrostaticsaided covalent self-assembly approach,w ed emonstrate efficient encapsulation of medium to large proteins (HRP,44kDa and b-gal, 465 kDa) and antibodies (ca. 150 kDa). The designed polymeric nanoassemblies are shown to successfully traffic functional antibodies (anti-NPC and anti-pAkt) to cytosol to elicit their bioactivity towardsb inding intracellular protein epitopes and inducing apoptosis.

Interface Assembly of Polymer Networks on Metal–Organic Frameworks for the Engineering of Functional Nanoparticles

“…Cross-linking strategies provide opportunities to engineer polymer NPs with desirable responsiveness to stimuli, such as redox and pH. − PEG NPs via cross-linking 8-arm-PEG-NH 2 and 8-arm-PEG-CHO based on Schiff base reaction were stable at neutral pH while disassembled in acidic environments (pH 5.5) due to the presence of pH-responsive imide bonds (Figure a). In addition, redox-sensitive HA NPs were assembled based on the formation of disulfide bonds between HA-SH and 8-arm-PEG-TNB, which could be disassembled in the presence of glutathione (GSH).…”

Optimizing Conjugation Chemistry, Antibody Conjugation Site, and Surface Density in Antibody–Nanogel Conjugates (ANCs) for Cell-Specific Drug Delivery

“…To assess the experimental conjugation reaction efficiency, we evaluated the reaction progression by gel electrophoresis at different time points after conjugation. We have previously used this method to evaluate protein encapsulation efficiency. , This evaluation was performed on ANC with different degrees of conjugation based on the weight ratio of antibody-to-polymeric nanogel weight (see the detailed calculation in the Materials and Methods Section). As shown in Figure c,d, nearly 90% of the antibody was consumed in the case of NG Tz and Ab TCO .…”

Antibody Polymer Conjugates (APCs) for Active Targeted Therapeutic Delivery

“…Following the DAR calculation of APCs, we evaluated the binding interactions using a flow assay in HER2+ SKOV3 cells. 27 To evaluate the possible loss in the targeting ability of the antibody due to the polymer conjugation, we first used a fluorophore-tagged secondary antibody that binds to the Fc domain of the primary antibody on the cell surface (Figure 3a). The specificity in binding would then be discerned from the fluorescence intensity differences achieved from the anti-HER2 Ab and the control IgG Ab.…”