title: Sustained cytoplasmic delivery and anti-viral effect of PLGA nanoparticles carrying a nucleic acid-hydrolyzing monoclonal antibody.
abstract: OBJECTIVE
  Cytoplasmic delivery of a monoclonal antibody (mAb) with nucleic acid-hydrolyzing activity (3D8 scFv) using poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs) was investigated for persistent anti-viral effect.
  
  
  METHODS
  3D8 scFv-loaded PLGA (3D8-PLGA) NPs were prepared via a double emulsion method that was previously optimized. Flow cytometry and confocal microscopy was carried out to confirm the cellular uptake and cytoplasmic localization. immunochemical and fluorescence resonance energy transfer (FRET) assays tested the cytoplasmic release and hydrolyzing effect of 3D8 scFv, respectively. Anti-viral activity test was performed using MTT assay with vesicular stomatitis virus (VSV)-infected HeLa cells.
  
  
  RESULTS
  3D8-PLGA NPs were much more effectively taken into cells in dose- and time-dependent manner and localized in the cytosolic region, compared to free 3D8 scFv. 3D8 scFv was released and hydrolyzed RNAs in the cytoplasm, exhibiting the maxima at a period of time (12-24 h). Anti-viral activity test revealed that 3D8-PLGA NP has dose- and time-dependent anti-viral effect and the maximum effect at the dose of 2 mg/ml and the incubation of 3 days.
  
  
  CONCLUSIONS
  Cytoplasmic delivery of 3D8 scFv via PLGA NPs could enhance the viability of infected cells in sustained manner due to preserved activity, much improved cellular uptake and sustained release.
authors.affiliation: Department of Molecular Science and Technology, Ajou University, San 5 Wonchon-dong, Yeongtong-Gu, Suwon, 443-749, Republic of Korea.
authors.forename: Yoon Ki
authors.initials: YK
authors.lastname: Joung
authors.forename: Sejin
authors.initials: S
authors.lastname: Son
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Jang
authors.forename: Myung Hee
authors.initials: MH
authors.lastname: Kwon
authors.forename: Ki Dong
authors.initials: KD
authors.lastname: Park
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000737:chemistry; D000998:Antiviral Agents / Q000008:administration & dosage* / Q000737:chemistry; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003600:Cytosol / Q000378:metabolism*; D003692:Delayed-Action Preparations; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods; D004655:Emulsions / Q000008:administration & dosage / Q000737:chemistry; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D019344:Lactic Acid / Q000008:administration & dosage* / Q000737:chemistry; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D009696:Nucleic Acids / Q000378:metabolism*; D011100:Polyglycolic Acid / Q000008:administration & dosage* / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012313:RNA / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D003692:Delayed-Action Preparations; D004337:Drug Carriers; D004655:Emulsions; D009696:Nucleic Acids; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D012313:RNA