Sustained Cytoplasmic Delivery and Anti-viral Effect of PLGA Nanoparticles Carrying a Nucleic Acid-Hydrolyzing Monoclonal Antibody

• Published in: Pharmaceutical research 2012-04, Vol.29 (4), p.932-942
• Main Authors: Joung, Yoon Ki, Son, Sejin, Jang, Ji Young, Kwon, Myung Hee, Park, Ki Dong
• Format: Article
• Language: English
• Subjects: Acids; Animals; Antibodies, Monoclonal - administration & dosage; Antibodies, Monoclonal - chemistry; Antiviral agents; Antiviral Agents - administration & dosage; Antiviral Agents - chemistry; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Cell Line, Tumor; Cell Survival - drug effects; Confocal microscopy; Controlled release; Cytoplasm; Cytosol - metabolism; Delayed-Action Preparations; Drug Carriers - administration & dosage; Drug Carriers - chemistry; Drug Delivery Systems - methods; Emulsions - administration & dosage; Emulsions - chemistry; Flow cytometry; fluorescence resonance energy transfer; General pharmacology; HCT116 Cells; HeLa Cells; Humans; Hydrolysis; Lactic Acid - administration & dosage; Lactic Acid - chemistry; Medical Law; Medical sciences; Mice; Monoclonal antibodies; Nanoparticles; Nanoparticles - administration & dosage; Nanoparticles - chemistry; Nucleic Acids - metabolism; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Pharmacology/Toxicology; Pharmacy; Polyglycolic Acid - administration & dosage; Polyglycolic Acid - chemistry; polylactide-co-glycolide; Research Paper; RNA - metabolism; Vesicular stomatitis virus
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-011-0633-0

ABSTRACT Purpose 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.