Direct Delivery of Functional Proteins and Enzymes to the Cytosol Using Nanoparticle-Stabilized Nanocapsules

Abstract

Intracellular protein delivery is an important tool for both therapeutic and fundamental applications. Effective protein delivery faces two major challenges: efficient cellular uptake and avoiding endosomal sequestration. We report here a general strategy for direct delivery of functional proteins to the cytosol using nanoparticle-stabilized capsules (NPSCs). These NPSCs are formed and stabilized through supramolecular interactions between the nanoparticle, the protein cargo, and the fatty acid capsule interior. The NPSCs are ~130 nm in diameter and feature low toxicity and excellent stability in serum. The effectiveness of these NPSCs as therapeutic protein carriers was demonstrated through the delivery of fully functional caspase-3 to HeLa cells with concomitant apoptosis. Analogous delivery of green fluorescent protein (GFP) confirmed cytosolic delivery as well as intracellular targeting of the delivered protein, demonstrating the utility of the system for both therapeutic and imaging applications.

Review of Applications and Environmental Fate of Gold Nanoparticles

“…Taking their drug delivery capability to a step higher, Tang et al, made GNPs-stabilized capsules using an oil core stabilized by amino acids conjugated with cationic GNPs. These capsules are capable of fusing directly with the cell membrane and have been found to be an efficient protein delivery system either into the cytosol or even more specifically to the nucleus [15]. Gene delivery can also be carried out through GNPs due to their excellent biocompatibility [16].…”

Nanoparticle–Dendrimer Hybrid Nanocapsules for Therapeutic Delivery

“…Arginine functionalized gold nanoparticles were synthesized as previously reported [10]. In a typical reaction, 10 mg of 1-pentanethiol protected gold nanoparticles was dissolved in 10 ml distilled DCM and purged with argon for 10 min.…”

“…We followed similar procedures to fabricate the NPSCs [10]. Briefly, 2.0 μl of linoleic acid was emulsified in 1 ml of arginine-functionalized nanoparticles (1.0 μM) in a phosphate buffer solution (5.0 mM, pH 7.4) using an amalgamator (speed 5000 rpm for 200 s).…”

“…NPSCs were initially stabilized through supramolecular interactions between the outer nanoparticle shell and the inner complementary hydrophobic 'oil' components. The NPSCs were further stabilized on the outer surface through lateral interactions between nanoparticles and complementarily charged biomolecules [10,11]. Using this strategy, we have been able to produce nanocapsules capable of effective delivery of paclitaxel (PTX), a small molecule chemotherapy drug with limited clinical applications due to its poor aqueous solubility [12].…”

Highly Versatile Polyelectrolyte Complexes for Improving the Enzyme Replacement Therapy of Lysosomal Storage Disorders

“…4 Within the context of protein delivery systems, stimulitriggered release is an appealing and promising approach, especially with the use of physiological stimuli like the pH drop within endo-lysosomal compartments. 5,6 In addition, addressing clinically relevant end points by targeted delivery has emerged as an approach to maximize the efficiency of a therapeutic macromolecule by strictly 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 5 localizing its pharmacological activity through controlled biodistribution.…”