論文 - 詳細
| RRC ID | 44932 |
|---|---|
| 著者 | Deo VK, Yui M, Alam J, Yamazaki M, Kato T, Park EY. |
| タイトル | A model for targeting colon carcinoma cells using single-chain variable fragments anchored on virus-like particles via glycosyl phosphatidylinositol anchor. |
| ジャーナル | Pharm Res |
| Abstract |
PURPOSE:VLPs displaying tumor targeting single-chain variable fragments (VLP-rscFvs) which targets tumor-associated glycoprotein-72 (TAG-72) marker protein have a potential for immunotherapy against colon carcinoma tumors. In this study, scFvs anchored on VLPs using glycosylphosphatidylinositol (GPI) were prepared to target colon carcinoma spheroids in vitro. METHODS:VLPs-rscFvs were produced by co-injecting two types of Bombyx mori nucleopolyhedrovirus (BmNPV) bacmids, encoding RSV-gag and rscFvs cDNA into silkworm larvae. Large unilamellar vesicles (LUVs) of 100 nm in diameter were made using 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and packaged with Sulforhodamine B (SRB). LUV-SRB was used to associate with VLP-rscFvs assisted by GP64 present on VLP-rscFvs to produce VLP-rscFv associated SRB (VLP-rscFvs-SRB) at pH 7.5. RESULTS:The antigenicity of the purified VLPs-rScFvs was confirmed by enzyme-linked immunosorbent assay (ELISA) using TAG-72 as antigen. LUV-SRB made of DOPC was used to associate with 100 μg of VLP-rscFvs to produce VLP-rscFv-SRB. Specific delivery and penetration of SRB up to 100 μm into the spheroids shows the potential of the new model. CONCLUSIONS:The current study demonstrated the display, expression and purification of VLP-rscFvs efficiently. As a test model VLP-rscFv-SRB were prepared which can be used for immunotherapy. rscFvs provide the specificity needed to target tumors and VLPs serve as carrier transporting the dye to target. |
| 巻・号 | 31(8) |
| ページ | 2166-77 |
| 公開日 | 2014-8-1 |
| DOI | 10.1007/s11095-014-1316-4 |
| PMID | 24570130 |
| MeSH | Animals Baculoviridae* / immunology Bombyx Cell Line, Tumor Colonic Neoplasms / drug therapy Drug Delivery Systems / methods* Glycosylphosphatidylinositols / administration & dosage* Glycosylphosphatidylinositols / immunology HEK293 Cells Hemolysis / drug effects Hemolysis / physiology Humans Single-Chain Antibodies / administration & dosage* Single-Chain Antibodies / immunology |
| IF | 3.242 |
| 引用数 | 8 |
| WOS 分野 | PHARMACOLOGY & PHARMACY CHEMISTRY, MULTIDISCIPLINARY |
| リソース情報 | |
| ヒト・動物細胞 | 293(RCB1637) |