An average synthesis yield of OCNs from graphite or graphene is limited to 8%. Although the previously reported OCN can be effectively used as a delivery reagent to bring matters into cells, there are many limitations on the OCN preparation. Our involvement in this area started from our preparation of the oxidized carbon nanospheres (OCNs) that possess excellent ability to bring macromolecules into cells 15, 16, 17. Apart from minimal toxicity, ideal reagents should possess simplicity during usages, and should be effective in delivering cargoes into cells without being destroyed by the commonly encountered endosome/lysosome pathway 13, 14. In addition to the use of cell penetrating peptides which require chemical coupling, and conventional liposomes which are unstable, a simple reagent that can effectively bring small peptides and big proteins into cells is, therefore, being needed 11, 12. Many of these applications require the transport of proteins into cells. In addition to many therapeutic applications, synthetic antibodies have been tailored as tools for various intracellular targets (intrabodies) 7, and have been successfully used for misfolded protein recognition 8, sensing protein conformation 9, and in vivo homing 10. Various specific affinitive macromolecules including RNA/DNA aptamers, siRNA, peptides and proteins have been tested with positive results 4, 5, 6. Remarkable advances in an understanding of signaling networks of disease progression together with developments of affinitive macromolecules in the past two decades, have made the interfering of biomolecular networks one of the most exciting researches and therapeutic means 1, 2, 3. An ability of OCBs to deliver big functional/therapeutic proteins into cells should open doors for more protein drug investigations and new levels of antibody therapies and biological studies. Delivery of human monoclonal antibodies (HuMAbs, 150 kDa) with specific affinity towards dengue viruses (DENV) into DENV-infected Vero cells by OCBs results in HuMAbs distribution all over cells’ interior and effective viral neutralization. Experiments with cell-sized liposomes indicate that OCBs directly interact with phospholipids and induce membrane leakages. Here we show that the relatively non-toxic and non-immunogenic oxidized carbon black particles (OCBs) prepared from commercially available carbon black can deliver a 300 kDa protein directly into cells, without an involvement of a cellular endocytosis. A simple and effective reagent that can bring functional proteins into cells can increase efficacy and allow more investigations. Modulating biomolecular networks in cells with peptides and proteins has become a promising therapeutic strategy and effective biological tools.
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