RIP nanoparticle formulation CHITOSAN â TPP CHAINS ARE ANTI-EpCAM conjugated TARGETED THERAPY FOR BREAST CANCER
ABSTRACT: Ribosome Inactivating Protein MJ-C is a protein from Mirabilis jalapa L. and has been tested in breast cancer cell line and shown cytotoxic properties. However RIP MJ-C were unstable and easily degraded. Chitosan nanoparticles can enhance RIP MJ-C stability and non-toxic for human use, therefore making it suitable as a carrier for RIP MJ-C. Nanoparticles were also conjugated with anti-EpCAM antibody which acted as targeting molecule to deliver RIP MJ-C nanoparticles to the cancer cells. The goal of this research is to develop a chitosan-TPP nanoparticle as a suitable carrier for delivering RIP MJ-C to the breast cancer cells. RIP MJ-C chitosan nanoparticles were prepared by ionotropic gelation method using Tripolyphosphate (TPP) as cross-linker. Various concentration of chitosan and TPP were screened to find a stable nanoparticle. Optimum formula for RIP MJ-C nanoparticles were previously determined by stability, entrapment efficiency and particle size. Nanoparticles made with optimum formula then conjugated with anti-EpCAM antibody and used for cytotoxicity assay toward T47D and fibroblast cells. These nanoparticles were analysed with FTIR to detect antibody conjugation. Chitosan-TPP RIP MJ-C nanoparticle were made using 0,5% chitosan and 0,01% TPP, with 50 µg/mL RIP MJ-C. These nanoparticles has spherical shape, 151.6 ± 121.38 nm particle size, 26.55 ± 0.8 mV zeta potential, and PI of 0.48 ± 0.06. Cytotoxicity assay shown that nanoparticle-encapsulated RIP MJ-C have higher cytotoxicity compared to naked RIP MJ-C. However antibody-conjugated RIP MJ-C nanoparticles showed no increase in cytotoxicity, and FTIR results showed that antibody conjugation to nanoparticles were only observed in nanoparticles without RIP MJ-C. These results showed that anti-EpCAM conjugated RIP MJ-C nanoparticles were successfully improved RIP MJ-C potency, but antibody conjugation was unsuccessful
