title: Formulation and Cytotoxicity of Ribosome-Inactivating Protein Mirabilis Jalapa L. Nanoparticles Using Alginate-Low Viscosity Chitosan Conjugated with Anti-Epcam Antibodies in the T47D Breast Cancer Cell Line.
abstract: Ribosome-inactivating protein (RIP) from Mirabilis jalapa L. leaves has cytotoxic effects on breast cancer cell lines but is less toxic towards normal cells. However, it can easily be degraded after administration so it needs to be formulated into nanoparticles to increase its resistance to enzymatic degradation. The objectives of this study were to develop a protein extract of M. jalapa L. leaves (RIP-MJ) incorporated into nanoparticles conjugated with Anti-EpCAM antibodies, and to determine its cytotoxicity and selectivity in the T47D breast cancer cell line. RIP-MJ was extracted from red-flowered M. jalapa L. leaves. Nanoparticles were formulated based on polyelectrolyte complexation using low viscosity chitosan and alginate, then chemically conjugated with anti-EpCAM antibody using EDAC based on carbodiimide reaction. RIP-MJ nanoparticles were characterised for the particle size, polydispersity index, zeta potential, particle morphology, and entrapment efficiency. The cytotoxicity of RIP-MJ nanoparticles against T47D and Vero cells was then determined with MTT assay. The optimal formula of RIP-MJ nanoparticles was obtained at the concentration of RIP-MJ, low viscosity chitosan and alginate respectively 0.05%, 1%, and 0.4% (m/v). RIP-MJ nanoparticles are hexagonal with high entrapment efficiency of 98.6%, average size of 130.7 nm, polydispersity index of 0.380 and zeta potential +26.33 mV. The IC50 values of both anti-EpCAM-conjugated and non-conjugated RIP-MJ nanoparticles for T47D cells (13.3 and 14.9 μg/mL) were lower than for Vero cells (27.8 and 33.6 μg/mL). The IC50 values of conjugated and non- conjugated RIP-MJ for both cells were much lower than IC50 values of non-formulated RIP-MJ (>500 μg/mL).
authors.affiliation: Postgraduated School of Biotechnology, Universitas Gadjah Mada, Yogyakarta, Indonesia E-mail : sismindari@ugm.ac.id.
authors.forename: Psycha Anindya
authors.initials: PA
authors.lastname: Wicaksono
authors.forename: Sismindari
authors.initials: S
authors.lastname: Name
authors.forename: Ronny
authors.initials: R
authors.lastname: Martien
authors.forename: Hilda
authors.initials: H
authors.lastname: Ismail
mesh_terms: D000464:Alginates / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D048271:Chitosan / Q000737:chemistry; D000071858:Epithelial Cell Adhesion Molecule / Q000037:antagonists & inhibitors* / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D039282:Mirabilis / Q000737:chemistry*; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D010316:Particle Size; D010936:Plant Extracts / Q000494:pharmacology*; D018515:Plant Leaves / Q000737:chemistry; D054788:Ribosome Inactivating Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014783:Viscosity
chemical_list: D000464:Alginates; D000911:Antibodies, Monoclonal; D000071858:Epithelial Cell Adhesion Molecule; D006603:Hexuronic Acids; D010936:Plant Extracts; D020723:Glucuronic Acid; D048271:Chitosan; D054788:Ribosome Inactivating Proteins