Preparation method and application of vesicle formed by erythrocyte membrane encapsulating Newcastle disease virus

1 . A preparation method of a vesicle formed by an erythrocyte membrane encapsulating Newcastle disease virus, comprising:

mixing a compacted erythrocyte solution, a phosphate buffered saline (PBS) solution, and Newcastle disease virus (NDV);

centrifuging the resulting mixture of erythrocytes added with NDV; and

resuspending the centrifuged pellet of the resulting mixture in a PBS solution to obtain the vesicle formed by the erythrocyte membrane encapsulating Newcastle disease virus;

wherein the compacted erythrocyte solution is obtained by:

diluting anticoagulated whole blood with an equal volume of PBS solution, separating erythrocyte, and removing the supernatant by centrifugation; and

adding a PBS solution to wash, and removing a supernatant by centrifugation to obtain the compacted erythrocyte solution;

wherein step of centrifuging the resulting mixture of erythrocytes added with NDV comprises:

collecting the mixture of erythrocytes with NDV and add the mixture into a 50 mL centrifuge tube;

centrifuging at 1300 mm 40° C. for 8 min, and then centrifuging at 5000 mm 40° C. for 15 min;

transferring the supernatant to another centrifuge tube and centrifuging at 14000 g 4° C. for 2 min; and

transferring the supernatant to another centrifuge tube and centrifuging at 14000 g 4° C. for 1 h;

wherein the step of resuspending the centrifuged pellet of the resulting mixture in a PBS solution to obtain the vesicle formed by the erythrocyte membrane encapsulating Newcastle disease virus comprises:

discarding the supernatant, resuspending the centrifuged pellet of the resulting mixture in 1 mL of PBS, and centrifuging at 14000 g 4° C. for 30 min; and

discarding the supernatant, resuspending the centrifuged pellet of the resulting mixture in 1 mL of PBS, and storing at −20° C.

2 . The method according to claim 1 , wherein the step of mixing a compacted erythrocyte solution, a phosphate buffered saline (PBS) solution, and Newcastle disease virus (NDV), comprises:

adding 10 mL of PBS solution and 4 mL NDV to each mL of the compacted erythrocyte solution, mix well, observing for hemagglutination after 1 hour, and retaining the mixture of erythrocytes with NDV when hemagglutination is observed and no erythrocyte deposition occurs, wherein the hemagglutination potency of NDV is 2 7 .

3 . A vesicle formed by an erythrocyte membrane encapsulating Newcastle disease virus prepared by the method according to claim 1 .

4 . A vesicle formed by an erythrocyte membrane encapsulating Newcastle disease virus prepared by the method according to claim 2 .