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Hybrid nanovaccine with photothermal response and preparation method thereof

A nano-vaccine, photothermal response technology, applied in the field of hybrid nano-vaccine and its preparation, can solve problems such as the increase of multi-drug resistant strains, and achieve the effects of low production cost, improved immune effect, and simple and controllable process.

Active Publication Date: 2019-08-30
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the abuse of antibiotics, Staphylococcus aureus strains have different degrees of drug resistance, and multi-drug resistant strains have also greatly increased. Drug-resistant Staphylococcus aureus has become one of the pathogenic bacteria that are difficult to suppress with existing antibiotics.

Method used

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  • Hybrid nanovaccine with photothermal response and preparation method thereof
  • Hybrid nanovaccine with photothermal response and preparation method thereof
  • Hybrid nanovaccine with photothermal response and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Preparation of OMV: Pick a single colony of Staphylococcus aureus of S.aureus BW15 and shake the bacteria in a 1L Erlenmeyer flask containing 250mL of LB medium at 37°C to the logarithmic phase (OD600 is about 1). Centrifuge at 6000g for 20min to remove the precipitate. After the supernatant is filtered through a 0.45μm filter membrane, the filtrate is concentrated to 20-30mL with a 100kDa ultrafiltration tube, and then filtered through a 0.22μm filter membrane. The filtrate is ultracentrifuged at 150,000g, 3h, 4℃, Precipitation is OMV 15 , washed three times with PBS, 150,000g, 3h, 4°C, to remove impurities;

[0047] (2) Preparation of ICG / MSN: Prepare 1 mg / mL ICG mother solution and dilute to 12 μg / mL, take 1 mL of ICG diluent, add 20 mg MSN, stir at room temperature for 2 h in the dark, and separate IgG / MSN on a magnetic stand;

[0048] (3) Preparation of OMV / ICG / MSN: take the OMV in step (1) respectively 15 Mix with ICG / MSN in step (2), the mass ratio is 1.5:1...

Embodiment 2

[0051] MSN, ICG / MSN, OMV in the preparation process of embodiment 1 15 / ICG / MSN was characterized and observed by transmission electron microscope and particle size analyzer. Such as image 3 As shown in A, mesoporous silicon can be effectively loaded with ICG and can be successfully coated by outer membrane vesicles; image 3 As shown in B, it is MSN, ICG / MSN, OMV 15 The particle sizes of / ICG / MSN are 88.6nm, 90.365nm and 96.3nm respectively, and the polydispersity coefficient PDI is 0.20, 0.27 and 0.22 respectively; image 3 C is its surface potential, MSN is positively charged (+26.6mV), ICG is negatively charged, and the potential decreases (+1.51mV) after MSN is loaded with ICG, while OMV 15 Negatively charged (-20.9mV), so wrap OMV 15 Post-OMV 15 / ICG / MSN becomes negative potential (-15.9mV).

Embodiment 3

[0053] The hybrid nanovaccine prepared in Example 1 was immunized into mice by subcutaneous injection, three times on day 0, day 7, and day 14, with a dose of 0.5 μg OMV protein, and then irradiated with 808nm laser for 5 minutes. Serum samples of the immunized mice were collected on days -1, 7, 14, and 21, and the anti-Staphylococcus aureus IgG titer in the serum was determined, as Figure 4 As shown in A, the prepared OMV has a strong humoral immune effect on mice, and after further preparation into a hybrid nano-vaccine and laser irradiation, the humoral immune effect is further enhanced. On the 21st day, take the spleen of the immunized mouse, prepare a single cell suspension, collect 2 million cells into a 1.5mL centrifuge tube, centrifuge at 1600rpm for 5min, remove the supernatant, wash gently with 1mL PBS twice, and finally resuspend in 100μL PBS , add CD3 and CD8 antibodies, and incubate at 4°C in the dark for 30min. Subsequently, it was fixed with 4% paraformaldehyd...

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Abstract

The invention discloses a hybrid nanovaccine with photothermal response and a preparation method thereof. The hybrid nanovaccine comprises magnetic mesoporous silicon, indocyanine green and staphylococcus aureus outer membrane vesicles (OMV). The invention uses bacterial OMV as the vaccine and the delivery carrier; at the same time, laser irradiation can make the indocyanine green produce heat topromote lysosome escape; reactive oxygen species (ROS) produced by lysosome rupture can further activate proteasomes to promote the processing of antigens; and finally, synergistic effect can be obtained to improve the immune effect of vaccine. The invention belongs to the field of pharmaceutical preparations and the technical field of bio-medicines. The nanovaccine is simple and controllable in preparation process, low in economic cost, good in repeatability and capable of effectively preventing the infection of drug-resistant staphylococcus aureus, with high clinical application value.

Description

technical field [0001] The invention relates to the field of pharmaceutical preparations and the field of biomedicine technology, in particular to a photothermal responsive hybrid nano-vaccine and a preparation method thereof. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus) is widely distributed in the natural environment and is one of the common clinical pathogens. Food poisoning caused by ingestion of staphylococcal enterotoxin is one of the most common foodborne diseases in the world, which seriously endangers public health. Due to the abuse of antibiotics, Staphylococcus aureus strains have drug resistance to varying degrees, and multi-drug resistant strains have also increased greatly. Drug-resistant Staphylococcus aureus has become one of the pathogenic bacteria that are difficult to suppress with existing antibiotics. The emergence of drug-resistant Staphylococcus aureus has brought great challenges to related treatments, so a new antibact...

Claims

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Application Information

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IPC IPC(8): A61K39/085A61K41/00A61K9/127A61K47/04A61P31/04
CPCA61K39/085A61K41/0057A61K41/0052A61K9/127A61K47/02A61P31/04A61K2300/00Y02A50/30
Inventor 陈刚周昕白亚南
Owner YANGZHOU UNIV
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