Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes

• Main Authors: Shi, Jiahai (Contributor), Kundrat, Lenka (Contributor), Pishesha, Novalia (Contributor), Bilate, Angelina M. (Contributor), Theile, Christopher S. (Contributor), Maruyama, Takeshi (Contributor), Dougan, Stephanie K. (Contributor), Ploegh, Hidde (Contributor), Lodish, Harvey F. (Contributor)
• Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Biology (Contributor), Whitehead Institute for Biomedical Research (Contributor)
• Format: Article
• Language: English
• Published: National Academy of Sciences (U.S.), 2015-02-04T15:45:13Z.
• Subjects: Article
• Online Access: Get fulltext

We developed modified RBCs to serve as carriers for systemic delivery of a wide array of payloads. These RBCs contain modified proteins on their plasma membrane, which can be labeled in a sortase-catalyzed reaction under native conditions without inflicting damage to the target membrane or cell. Sortase accommodates a wide range of natural and synthetic payloads that allow modification of RBCs with substituents that cannot be encoded genetically. As proof of principle, we demonstrate site-specific conjugation of biotin to in vitro-differentiated mouse erythroblasts as well as to mature mouse RBCs. Thus modified, RBCs remain in the bloodstream for up to 28 d. A single domain antibody attached enzymatically to RBCs enables them to bind specifically to target cells that express the antibody target. We extend these experiments to human RBCs and demonstrate efficient sortase-mediated labeling of in vitro-differentiated human reticulocytes.
United States. Defense Advanced Research Projects Agency (Contract HR0011-12-2-0015)