In this project, we are determined to create potential oxygen carriers, transporting hemoglobin using the cargo loading of exosomes, for the development of artificial blood. PNCV plasmids have been transfected into HEK 293T cell line, and the successful translation of Vhb (Vitreoscilla hemoglobin) was proved by western blot results. Exosomes were extracted from the medium. SDS-PAGE results and TEM observation both yielded expected results, demonstrating the feasibility of this extraction. Attaching Vhb to exosomes didn’t affect either protein, which was again shown by TEM results. COD testing proved that the heme group was functioning successfully: better in the case of membrane anchored protein CD63 than that of WWtag3. Repeated testing, as well as comparison between control groups and fusion protein groups demonstrated the success of experiment: by concluding all experimental evidence, we see that an efficient oxygen carrier, composed of Vhb and exosome, has been produced. Although more experiments still have to be performed to demonstrate its behaviors in vivo, and more considerations for its practice need to be taken, we consider it an essential step in the whole process of developing artificial blood.

Key Prove:

Western blot results indicated production of Vhb proteins.

SDS-PAGE yielded correct protein bonds for exosomes. TEM visualized the correct shape and size.

TEM demonstrated that the fusion of Vhb didn’t affect the shape or concentration of exosomes. COD proved the potential function of fusion proteins to transport oxygen efficiently. The loading of Vhb into exosome has been successful.