title: A simplified method to attach antibodies on liposomes by biotin-streptavidin affinity for rapid and economical screening of targeted liposomes.
abstract: The biotin-Streptavidin (STREP) technique for attachment of monoclonal antibodies (mAbs) (or other ligand types) on liposome surface offers high attachment yield, however it is time consuming and expensive due to the number of steps used and the consumption of large quantities of STREP. Herein, a simplified, fast and economic technique, by incubating pre-mixed biotin-mAb/STREP with biotin-liposomes, at a 3:1:1 biotin-mAb/STREP/biotin-LIP ratio (mol/mol/mol) was evaluated. The physichochemical properties, final mAb attachment yield and targeting potential of liposomes decorated with an anti-transferrin receptor mAb (TfR-mAb), prepared by the simple method (SM) and the conventional method (CM), were compared. The vesicle uptake by hCMEC/D3 cells (known to overexpress TfR) were considered as a measure of liposome targeting capability. Results show that both targeted liposome types (SM and CM) have small size (mean diameters around 150 nm), low poly-dispersity (approx. 0.20) and similar mAb attachment yield (between 64-88%). However, the uptake of the SM-liposomes is slightly lower compared to CM-LIP (24-30% decrease), suggesting that the modulated conformation of mAbs on the liposome surface (triplets attached to one single STREP molecule) results in decreased targeting capability. Nevertheless, the simpler and faster one-step preparation procedure which has very high lipid recovery (> 95%) compared to the CM (50-60%) and 15-30 times lower consumption of STREP, may be a good alternative for initial screening of various mAbs as ligands for targeted liposomal or other nanotechnologies, during pre-clinical development.
authors.forename: Konstantina
authors.initials: K
authors.lastname: Papadia
authors.forename: Eleni
authors.initials: E
authors.lastname: Markoutsa
authors.forename: Sophia G
authors.initials: SG
authors.lastname: Antimisiaris
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D001710:Biotin / Q000737:chemistry* / Q000276:immunology; D003460:Crystallization / Q000379:methods*; D007118:Immunoassay / Q000379:methods*; D008081:Liposomes / Q000737:chemistry* / Q000276:immunology; D008422:Materials Testing; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D019809:Streptavidin / Q000737:chemistry* / Q000276:immunology
chemical_list: D000906:Antibodies; D008081:Liposomes; D001710:Biotin; D019809:Streptavidin