Nanomaterials are the foundations of Nanotechnology, which are measured in nanoscales, Carbon nanotubes are one of the interesting nanomaterials, studied for over 25 years because of their superlative properties such as high surface area, electrical and thermal conductivity, high biocompatibility, flexibility, resistance to corrosion and nanosize. According to research, carbon nanotubes are applied in sensing, water treatment, and drug delivery, mainly used to deliver the anticancer drugs. In our work, functionalization of multi walled carbon nanotubes done by covalent and non-covalent functionation methods, covalent functionalization showed better dispersing efficiency in aqueous medium and compatible with biological systems with damaging the crystal lattice of carbon nanotubes. Non covalent functionalization helps to derivatized with active compounds, surface adsorption or attachment of various molecules or antibodies, which subsequently helps in targeting the site and to produce therapeutic effects. Different formulations prepared by functionalized MWCNTs and multiple functionalization of MWCNTs done by binding the drug and antibodies to prepare functionalized MWCNTs 5-Fluorouracil complexes. The Cytotoxicity assay was carried out for the obtained new targeting formulations to analyze the effect of all the formulations on HCT116 cell line. The percentage death was determined based on the viability of the cells in the appropriate vehicle controls. In this study, we report the successful functionalization, binding of 5 Fluorouracil, antibodies to MWCNTs, and cells viability of all prepared formulations for the development of novel carbon based anticancer drug delivery system. Functionalized MWCNTs-5-Fluorouracil antibodies composite at concentration above 2.5 µg/mL exhibited ≥ 50% cytotoxicity post normalization with compound control to negate precipitation observed with the compound. All the formulations showed the precipitations indicating antitumor activity and biocompatibility.