US 12,648,990 B2 · Bacterial membrane vesicles, and separation and preparation system and method therefor

Abstract

The present invention belongs to the field of microbiology, and particularly relates to membrane vesicles (MVs) isolated from bacteria, and an isolation and preparation system and method for the membrane vesicles, and applications of the membrane vesicles. The bacteria of the present invention comprise Gram-positive bacteria and Gram-negative bacteria. The invention uses ionizing irradiation to irradiate bacteria, and isolates and purifies the produced membrane vesicles. The membrane vesicles prepared can be used as a vaccine, a vaccine adjuvant and/or a pharmaceutical carrier. In addition, the present invention provides a biological composition comprising the membrane vesicles and inactivated bacteria. In addition, the present invention also provides a preparation system, and isolation and purification system for bacterial membrane vesicles and the corresponding method. The membrane vesicles obtained by using the system and method have high yield, high purity and easy to be industrialized.

Claims9

Dependent claims

A method for preparing membrane vesicles of bacteria, wherein the bacteria comprise Pseudomonas aeruginosa or Staphylococcus aureus, and the method comprises the following steps:

Step 1) Culturing the bacteria to logarithmic growth phase;
Step 2) Collecting bacterial cells, resuspending the bacterial cells to obtain a bacterial solution, and irradiating the bacterial solution with ionizing irradiation to obtain irradiated bacterial solution, wherein the ionizing irradiation is X-rays, and irradiation dose is 500-1000 Gy;
Step 3) Collecting the irradiated bacterial solution, centrifuging the irradiated bacterial solution at 100-10000 g for 10-60 min, collecting supernatant 1, and filtering the supernatant 1 with a 0.3-0.5 μM filter to remove the bacterial cells and obtain filtered supernatant 1;
Step 4) Centrifuging the filtered supernatant 1 at 5000-25000 g for 10-100 min, collecting supernatant 2;
Step 5) Centrifuging the supernatant 2 at 5000-150000 g for 60-600 min to precipitate and obtain the membrane vesicles.

2 more elements

Dependencies are inferred from the claim text.

Bacterial membrane vesicles, and separation and preparation system and method therefor

Life of this patent

Bacterial membrane vesicles, and separation and preparation system and method therefor

Life of this patent

Abstract

Claims9