Abstract
1-ethyl-3-(3-dimethylaminopropyl) carbondiimide hydrochloride (EDC) is commonly usedas a crosslinker to help bind biomolecules, such as DNA plasmids, with nanostructures.However, EDC often remains, after a crosslink reaction, in the micro-aperture of thenanostructure, e.g., carbon nanotube. The remaining EDC shows positive green fluorescentsignals and makes a nanostructure with a strong cytotoxicity which induces cell death. Thetoxicity of EDC was confirmed on a breast cancer cell line (MCF-7) and two leukemiccell lines (THP-1 and KG-1). The MCF-7 cells mainly underwent necrosis aftertreatment with EDC, which was verified by fluorescein isothiocyanate (FITC)annexin V staining, video microscopy and scanning electronic microscopy (SEM). Ifthe EDC was not removed completely, the nanostructures with remaining EDCproduced a green fluorescent background that could interfere with flow cytometry(FACS) measurement and result in false information about GFP plasmid delivery.Effective methods to remove residual EDC on macromolecules were also developed.
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