Antibody-loading of biological nanocarrier vesicles derived from red-blood-cell membranes

Antibodies, disruptive potent therapeutic agents against pharmacological targets, face a barrier crossing immune-system and cellular-membranes. To overcome these, various strategies have been explored including shuttling via liposomes or bio-camouflaged nanoparticles. Here, we demonstrate the feasibility to load antibodies into exosome-mimetic nanovesicles derived from human red-blood-cell-membranes. The goat-anti-chicken antibodies are loaded into erythrocyte-membrane derived nanovesicles and their loading yields are characterized and compared with smaller dUTP-cargo. Applying dual-color coincident fluorescence burst methodology, the loading yield of nanocarriers is profiled at single-vesicle level overcoming their size-heterogeneity and achieving a maximum of 38-41% antibody-loading yield at peak radius of 52 nm. The average of 14 % yield and more than two antibodies per vesicle is estimated, comparable to those of dUTP-loaded nanovesicles after additional purification through exosome-spin-column. These results suggest a promising route for enhancing biodistribution and intracellular accessibility for therapeutic antibodies using novel, biocompatible, and low-immunogenicity nanocarriers, suitable for large-scale pharmacological applications.