Preparation and Characterization of Paclitaxel-loaded PLGA nanoparticles coated with cationic SM5-1 single-chain antibody.

The purpose of this study was to develop paclitaxel-loaded poly(lactide-co-glycolide) (PLGA) nanoparticles coated with cationic SM5-1 single-chain antibody (scFv) containing a polylysine (SMFv-polylys). SM5-1 scFv (SMFv) is derived from SM5-1 monoclonal antibody, which binds to a 230 kDa membrane protein specifically expressed on melanoma, hepatocellular carcinoma and breast cancer cells. SMFv-polylys was expressed in Escherichia coli and purified by cation-exchange chromatography. Purified SMFv-polylys was fixed to paclitaxel-loaded PLGA nanoparticles to form paclitaxel-loaded PLGA nanoparticles coated with SMFv-polylys (Ptx-NP-S). Ptx-NP-S was shown to retain the specific antigen-binding affinity of SMFv-polylys to SM5-1 binding protein-positive Ch-hep-3 cells. Finally, the cytotoxicity of Ptx-NP-S was evaluated by a non-radioactive cell proliferation assay. It was demonstrated that Ptx-NP-S had significantly enhanced in vitro cytotoxicity against Ch-hep-3 cells as compared with non-targeted paclitaxel-loaded PLGA nanoparticles. In conclusion, our results suggest that cationic SMFv-polylys has been successfully generated and may be used as targeted ligand for preparing cancer-targeted nanoparticles.

[1]  H. Drobecq,et al.  Expression and purification of recombinant vascular endothelial-statin. , 2006, Protein expression and purification.

[2]  Y. Antonenko,et al.  Grafting of polylysine with polyethylenoxide prevents demixing of O-pyromellitylgramicidin in lipid membranes. , 2006, Biochimica et biophysica acta.

[3]  Shubiao Zhang,et al.  Toxicity of cationic lipids and cationic polymers in gene delivery. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[4]  Hao Wang,et al.  Preparation and characterization of fusion protein truncated Pseudomonas Exotoxin A (PE38KDEL) in Escherichia coli. , 2005, Protein expression and purification.

[5]  Weizhu Qian,et al.  Construction and characterization of a high-affinity humanized SM5-1 monoclonal antibody. , 2007, Biochemical and biophysical research communications.

[6]  Xin Lu,et al.  The cytotoxic effect of E1B 55-kDa mutant adenovirus on human hepatocellular carcinoma cell lines , 2001, Cancer Gene Therapy.

[7]  Richard Perron,et al.  Eu3+-doped Gd2O3 nanoparticles as reporters for optical detection and visualization of antibodies patterned by microcontact printing , 2006, Analytical and bioanalytical chemistry.

[8]  H. Ghandehari,et al.  Nanocarriers for nuclear imaging and radiotherapy of cancer. , 2006, Current pharmaceutical design.

[9]  Jayanth Panyam,et al.  Biodegradable nanoparticles for drug and gene delivery to cells and tissue. , 2003, Advanced drug delivery reviews.

[10]  Ya-jun Guo,et al.  The monoclonal antibody SM5-1 recognizes a fibronectin variant which is widely expressed in melanoma , 2006, BMC Cancer.

[11]  J. Richie,et al.  Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[12]  S. Nathan,et al.  Bacterial expression of the scFv fragment of a recombinant antibody specific for Burkholderia pseudomallei exotoxin. , 2003, Journal of biochemistry and molecular biology.

[13]  Yanjun Liu,et al.  SM5-1: a new monoclonal antibody which is highly sensitive and specific for melanocytic lesions , 2000, Archives of Dermatological Research.

[14]  T. Allen.  Ligand-targeted therapeutics in anticancer therapy , 2002, Nature Reviews Cancer.

[15]  S. Simões,et al.  Paclitaxel-loaded PLGA nanoparticles: preparation, physicochemical characterization and in vitro anti-tumoral activity. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[16]  Y. Li,et al.  Expression vectors for human-mouse chimeric antibodies. , 2005, Journal of biochemistry and molecular biology.

[17]  Roger Le Grand,et al.  Surfactant-free anionic PLA nanoparticles coated with HIV-1 p24 protein induced enhanced cellular and humoral immune responses in various animal models. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[18]  S. Panyim,et al.  Anti-CHH antibody causes impaired hyperglycemia in Penaeus monodon. , 2006, Journal of biochemistry and molecular biology.

[19]  P. Carter,et al.  Improving the efficacy of antibody-based cancer therapies , 2001, Nature Reviews Cancer.

[20]  Manmohan J. Singh,et al.  Encapsulation of the immune potentiators MPL and RC529 in PLG microparticles enhances their potency. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[21]  Tae Gwan Park,et al.  Target-specific cellular uptake of PLGA nanoparticles coated with poly(L-lysine)-poly(ethylene glycol)-folate conjugate. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[22]  Manmohan J. Singh,et al.  An investigation of the factors controlling the adsorption of protein antigens to anionic PLG microparticles. , 2005, Journal of pharmaceutical sciences.

[23]  G. Katzenmeier,et al.  High level of soluble expression in Escherichia coli and characterisation of the cloned Bacillus thuringiensis Cry4Ba domain III fragment. , 2007, Journal of biochemistry and molecular biology.

[24]  Hao Wang,et al.  Differential Expression of MART-1, Tyrosinase, and SM5-1 in Primary and Metastatic Melanoma , 2005, The American Journal of dermatopathology.

[25]  J. Panyam,et al.  Susceptibility of nanoparticle-encapsulated paclitaxel to P-glycoprotein-mediated drug efflux. , 2006, International journal of pharmaceutics.

[26]  H. Sah.  A new strategy to determine the actual protein content of poly(lactide-co-glycolide) microspheres. , 1997, Journal of pharmaceutical sciences.

[27]  Robert Gurny,et al.  Current methods for attaching targeting ligands to liposomes and nanoparticles. , 2004, Journal of pharmaceutical sciences.

[28]  L. Lim,et al.  Preparation and in vitro anticancer activity of wheat germ agglutinin (WGA)-conjugated PLGA nanoparticles loaded with paclitaxel and isopropyl myristate. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[29]  A. Moshfeghi,et al.  Micro- and nanoparticulates. , 2005, Advanced drug delivery reviews.

[30]  Hao Wang,et al.  Treatment of hepatocellular carcinoma in a mouse xenograft model with an immunotoxin which is engineered to eliminate vascular leak syndrome , 2007, Cancer Immunology, Immunotherapy.

[31]  K. Mizuno,et al.  Monoclonal Antibody , 2020, Definitions.