FORMULATION AND TEST cytotoxic ribosome inactivating protein nanoparticles Mirabilis jalapa.L (RIP MJ-C) CHITOSAN USING SHORT CHAIN AND ANTI-conjugated pectin methylation EpCAM LOW ON BREAST CANCER CELL T47D
ABSTRACT: RIP MJ-C in Mirabilis jalapa .L plant is obtained through the purification process by using CM-Sepharose CL-6B stationary phase. To protect the RIP MJ-C from the effects of degradation, a nanoparticle system by using chitosan polymer short chain and low methylation pectin which conjugated with anti-EpCAM antibody 9C4 to improve selectivity and effectiveness against breast cancer cells T47D. The extraction of M. jalapa L leaves are conducted with liquid mixing media technique, called pH 7.2 phosphate buffer. The extraction of M. jalapa L leaves are purified by using ion exchange chromatography method, the result is RIP MJ-C. Before and after RIP MJ-C is being electrophoresis lyophilized to examine its activities. The optimum formula nanoparticles determined by using factorial design 22 method with Design Expert software version 7.1.5. Formula nanoparticles RIP MJ-C using chitosan polymer short chain and low methylation pectin which conjugated with anti-EpCAM is tested its cytotoxicity effects by MTT assay method. The extraction result of 1718,5 g M. jalapa L leaves produce 1,656 g of RIP MJ-C. Based on the optimization, optimum formula is gained from a combination of short chain chitosan 0,06% and 0,02%. Nanoparticles characteristic which was produced with particle size of 54,43± 10,14 nm, polydispersity index of 0,514 ± 0,10, and + zeta potential of 93,59± 6,90 mV. Electrophoresis results showed that before and after RIP MJ-C was being lyophilized has an activity to cut supercoil DNA to become circular nick and linear shapes. The result of cytotoxic test shows the data in percentage indicates the highest percentage of the death cell (43,20%) in the nanoparticles formula conjugated with 16,67 µg/mLconcentration of anti-EpCAM 9C4. From the results of this study, it can be concluded that the RIP can be encapsulated in a nanoparticle system, the nanoparticles were conjugated with anti-EpCAM can enhance cytotoxic effects.
