Fe_(3)O_(4)@SiO_(2)-Protein A-oHSV/CD63 Ab for Capturing Virus and Exosomes

Protein A modified magnetic spheres(Fe_(3)O_(4)@SiO_(2)-Protein A)with 22.7 emu/g saturation magnetization were prepared and characterized.These spheres had a diameter of 10 nm and was stable up to 229.2℃,and can bond 19.694 mg/g of Protein A.The Fe_(3)O_(4)@SiO_(2)-Protein A was firstly combined with oHSV-BJ-2-A antibody to capture oHSV from disease venom,and the amount of binding virus was 1.92×10^(7)to 3.44×10^(7)PFU/g.Fe_(3)O_(4)@SiO_(2)-Protein A was modified with CD63 antibody,which was further fixed using dissuccinimide octylate(DSS)as a crosslinking agent.The prepared Fe_(3)O_(4)@SiO_(2)-Protein A-CD63 Ab was useful in isolating and enriching exosomes from cell supernatant.Compared with commercial kits,Fe_(3)O_(4)@SiO_(2)-Protein A-CD63 Ab demonstrated a better purification effect,which was successfully monitored using capillary electrophoresis.The total content of exosomes protein extracted using Fe_(3)O_(4)@SiO_(2)-Protein A-CD63 was 3.4 mg/g,which was obviously higher than that reported in some studies.In addition,Fe_(3)O_(4)@SiO_(2)-Protein A-CD63 Ab also showed its repeatability through bonding and elution of 5 cycles,which was effective in cost saving.These indicated Fe_(3)O_(4)@SiO_(2)-Protein A-CD63 Ab had the potential for large-scale purification of exosomes in practical applications.