Single chain variable fragment fused to maltose binding protein: a modular nanocarrier platform for the targeted delivery of antitumorals

• Published in: Biomaterials science 2021-03, Vol.9 (5), p.1728-1738
• Main Authors: Reche-Perez, Francisco J, Plesselova, Simona, De los Reyes-Berbel, Eduardo, Ortega-Muñoz, Mariano, Lopez-Jaramillo, Francisco J, Hernandez-Mateo, Fernando, Santoyo-Gonzalez, Francisco, Salto-Gonzalez, Rafael, Giron-Gonzalez, Maria D
• Format: Article
• Language: English
• Subjects: Adapters; Animals; Antibodies, Monoclonal; Chains; Chemical synthesis; Coordination compounds; Covalence; Cyclodextrins; Doxorubicin; Fragments; Growth factors; Maltose; Maltose-Binding Proteins - genetics; Mice; Modularity; Monoclonal antibodies; Oligonucleotides; Optimization; Proteins; Reagents; Single-Chain Antibodies - genetics; Xenotransplantation
• ISSN: 2047-4830; 2047-4849
• DOI: 10.1039/d0bm01903h

The use of the specific binding properties of monoclonal antibody fragments such as single-chain variable fragments (ScFv) for the selective delivery of antitumor therapeutics for cancer cells is attractive due to their smaller size, low immunogenicity, and low-cost production. Although covalent strategies for the preparation of such ScFv-based therapeutic conjugates are prevalent, this approach is not straightforward, as it requires prior chemical activation and/or modification of both the ScFv and the therapeutics for the application of robust chemistries. A non-covalent alternative based on ScFv fused to maltose-binding protein (MBP) acting as a binding adapter is proposed for active targeted delivery. MBP-ScFv proves to be a valuable modular platform to synergistically bind maltose-derivatized therapeutic cargos through the MBP, while preserving the targeting competences provided by the ScFv. The methodology has been tested by using a mutated maltose-binding protein (MBP I334W) with an enhanced affinity toward maltose and an ScFv coding sequence toward the human epidermal growth factor receptor 2 (HER2). Non-covalent binding complexes of the resulting MBP-ScFv fusion protein with diverse maltosylated therapeutic cargos (a near-infrared dye, a maltosylated supramolecular β-cyclodextrin container for doxorubicin, and non-viral polyplex gene vector) were easily prepared and characterized. In vitro and in vivo assays using cell lines that express or not the HER2 epitope, and mice xenografts of HER2 expressing cells demonstrated the capability and versatility of MBP-ScFv for diagnosis, imaging, and drug and plasmid active targeted tumor delivery. Remarkably, the modularity of the MBP-ScFv platform allows the flexible interchange of both the cargos and the coding sequence for the ScFv, allowing ad hoc solutions in targeting delivery without any further optimization since the MBP acts as a pivotal element. A modular platform for targeted delivery was based on a single chain variable (ScFv) fragment fused to maltose-binding protein (MBP). Using different maltosylated ligands it is likely to target, transport drugs, or deliver genes to specific cells.