Monoclonal antibody covalently coupled to liposomes: specific targeting to cells.
We have evaluated optimal conditions for coupling monoclonal antibody to small unilamellar liposomes. Coupling of an IgG2 alpha monoclonal anti-beta 2-microglobulin antibody, which reacts with human cells, was examined in detail. Liposomes were composed of dipalmitoyl lecithin and cholesterol, and variable quantities of phosphatidylethanolamine substituted with the heterobifunctional cross-linking reagent N-hydroxysuccinimidyl 3-(2-pyridyldithio) propionate (SPDP). They were reacted with antibody derivatized with the same reagent at a 5- to 20-fold molar excess, and activated by mild reduction. This degree of SPDP modification had no effect on the capacity of the antibody to bind to its target antigen. More than 40% of antibody could be reproducibly bound to liposomes, resulting in the coupling of from 1 to 10 antibody molecules per liposome (mean diameter:580 A). The coupling reaction did not lead to loss of carboxyfluorescein encapsulated within liposomes. At least 80% of liposomes carried nondenatured antibody, as confirmed by precipitation of liposomes and encapsulated carboxyfluorescein by Staphylococcus aureus, strain Cowan I. The liposome-coupled antibody retained its immunological specificity: only cells expressing human beta 2-microglobulin bound liposomes in vitro, and the binding was inhibited by the free antibody in solution. Results with antibodies of different antigens specificity confirm that the technique can be generally applied.
D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes; D002460:Cell Line; D002784:Cholesterol; D006801:Humans; D007700:Kinetics; D007945:Leukemia, Lymphoid; D008081:Liposomes; D011663:Pulmonary Surfactants; D011863:Radioimmunoassay; D001613:beta 2-Microglobulin / Q000276:immunology
D000911:Antibodies, Monoclonal; D008081:Liposomes; D011663:Pulmonary Surfactants; D001613:beta 2-Microglobulin; D002784:Cholesterol