Silencing red blood cell recognition toward Anti-A antibody by means of polyelectrolyte layer-by-layer assembly in a two-dimensional model system.

Silencing the antigenic response of red blood cells (RBCs) is a prerequisite toward the development of universal blood transfusion. Using a two-dimensional (2D) model whereby nonfixed RBCs are adsorbed on a human fibronectin (HFN)-coated surface, we demonstrate that the layer-by-layer (LbL) assembly technique of biocompatible polyelectrolytes can be employed to achieve the immunocamouflage of RBCs against the Anti-A antibody while maintaining the integrity and viability of the cells. The multilayered film consisted of a protecting shell (P-shell), containing five bilayers of chitosan-graft-phosphorylcholine (CH-PC) and sodium hyaluronate (HA), covered by a camouflage shell (C-shell) made up of five bilayers of poly-(L-lysine)-graft-poly(ethylene glycol) (PLL-PEG) and alginate (AL). Control experiments in which RBCs were coated by (CH-PC/HA)(10) bilayers indicated that the two polyelectrolytes alone did not prevent immunorecognition. The LbL film formation on RBCs and model substrates was monitored by quartz crystal microbalance with dissipation factor (QCM-D) and analyzed through zeta-potential measurements, atomic force microscopy (AFM), and optical microscopy. Antibody interaction with the coated RBCs was investigated by QCM-D, fluorescence microscopy, and hemolysis assays. Results from these measurements demonstrated that the hybrid LbL system built-up with different sets of polyelectrolytes was able to protect the RBCs from hemolysis and recognition by the Anti-A antibody.