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Manganese nano adjuvant as well as preparation method and application thereof

A nano and adjuvant technology, applied in the field of biomedicine, can solve the problems of the immune enhancement effect to be improved

Pending Publication Date: 2021-05-14
THE GBA NAT INST FOR NANOTECHNOLOGY INNOVATION +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immune enhancement effect of pure divalent manganese adjuvant or tetravalent manganese adjuvant needs to be improved

Method used

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  • Manganese nano adjuvant as well as preparation method and application thereof
  • Manganese nano adjuvant as well as preparation method and application thereof
  • Manganese nano adjuvant as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The steps of preparing manganese nano-adjuvant are as follows ( figure 1 ):

[0048] A) Add 5mL manganese chloride aqueous solution with a concentration of 0.1mol / L to 10mL bovine serum albumin (BSA) solution with a concentration of 10mg / mL, stir and mix to obtain a composite solution.

[0049] B) Add 0.1 mL of NaOH aqueous solution with a concentration of 2 mol / L to the composite solution in step A, continue to stir at 34 °C and 900 rpm for 2 hours, dialyze, purify and freeze-dry to obtain the manganese nano-adjuvant (MnARK) product.

[0050] see figure 2 . figure 2 Transmission electron microscope image and X-ray diffraction spectrum of the manganese nano-adjuvant prepared in Example 1 of the present invention.

[0051] Depend on figure 2 It can be seen that the average particle size of the manganese nano-adjuvant prepared in Example 1 of the present invention is 9.77 nanometers, and has good dispersion and uniform size; XRD data shows that the manganese nano-adj...

Embodiment 2

[0053] The steps of preparing manganese nano adjuvant are as follows:

[0054] A) Add 10mL of manganese nitrate aqueous solution with a concentration of 0.2mol / L to 5mL of 50mg / mL human serum albumin (HSA) solution, stir and mix to obtain a complex solution.

[0055] B) Add 0.5mL of 0.1mol / L NaOH aqueous solution to the composite solution in step A, continue to stir at 30°C and 700rpm for 5 hours, dialyze, purify and freeze-dry to obtain the manganese nano adjuvant product.

[0056] see image 3 . image 3 TEM image of the manganese nano-adjuvant prepared for Example 2 of the present invention.

[0057] Depend on image 3 It can be seen that the average particle size of the manganese nano-adjuvant prepared in Example 2 of the present invention is 28.43 nm, and has good dispersion and uniform size.

Embodiment 3

[0059] The biotoxicity and immune cell activation test of the manganese nano adjuvant (MnARK) obtained in Example 1.

[0060] A) Viability assay: DC2.4 cells were seeded in 96-well plates and incubated with different concentrations (0 to 0.5 mmol / L) of manganese chloride or MnARK for 24 hours. Cell viability was assessed using the CCK-8 kit.

[0061] B) Flow cytometry analysis experiment: DC2.4 cells were divided into 3×10 cells per well 5 The density of cells was seeded in a 6-well plate and cultured for 12 hours, then the cells were treated with manganese chloride or MnARK, and after 24 hours of incubation, the cells were collected and treated with anti-CD11c, anti-CD80 and anti-CD86 (all purchased from TONBO Biosciences) Stained for flow cytometry analysis (BD Accuri C6, BD, USA).

[0062] See Figure 4 . Figure 4 The biological toxicity of the manganese nano-adjuvant (MnARK), the internalization of the nano-adjuvant by DC2.4 cells and the promotion of DC maturation in...

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Abstract

The invention relates to the technical field of biological medicines and vaccines, in particular to a manganese nano adjuvant as well as a preparation method and application thereof. The manganese nano adjuvant comprises manganous-manganic oxide nano particles and template molecules wrapping the manganous-manganic oxide nano particles, the molar ratio of the template molecules to manganese is 1: (10-1000), and the template molecules comprise template protein and fragments or polypeptides thereof. The manganese nano adjuvant provided by the invention can effectively carry an immune antigen, and can obtain a more excellent immunotherapy effect when the antigen carrying amount is relatively small and the injection usage amount is relatively low; the immune antigen can be effectively delivered to lymph node tissues, cell internalization of the immune antigen is greatly enhanced, and immune cells are activated very efficiently.

Description

technical field [0001] The invention relates to the fields of biomedical technology and vaccine technology, in particular to a manganese nano adjuvant, its preparation method and application. Background technique [0002] In addition to optimizing immune antigens, using appropriate nano-adjuvants to improve the immunogenicity of new crown antigens, reduce the number of vaccinations and antigen doses, and induce effective neutralizing antibodies and cell-mediated immune responses is an effective way to prevent the new crown from continuing to be a pandemic. One of the potential coping strategies. [0003] Aluminum adjuvant is recognized as a safe adjuvant. Compared with free antigen, aluminum adjuvant can promote better immune response to antigen, but at the same time, the lack of cellular immunity is a natural defect of alum-prepared vaccines. Currently, the design and construction of vaccine systems to achieve satisfactory antigen delivery (lymph node (LN) targeting and ef...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K47/42A61K39/00A61K39/215A61P31/14
CPCA61K39/39A61K47/42A61K39/00A61K39/12A61P31/14A61K2039/55505A61K2039/572A61K2039/575A61K2039/54C12N2770/20034
Inventor 陈春英王亚玲刘野郭梦雨陈曦胡栩豪赵宇亮
Owner THE GBA NAT INST FOR NANOTECHNOLOGY INNOVATION
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