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Nano coronavirus recombinant vaccine taking graphene oxide as carrier

A coronavirus and graphene technology, applied in the fields of nanomaterials and biomedicine, can solve the problems of vaccine development failure, inability to produce immune memory, vaccine approval and marketing, etc., and achieve strong immunogenicity

Pending Publication Date: 2021-01-15
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, more studies on SARS and MERS vaccines are mainly focused on viral vector vaccines and subunit vaccines. A large number of studies have shown that the difficulty of SARS and MERS is that they cannot stimulate the production of long-term memory B cells. In cured SARS and MERS patients, long-term memory cells only It can last for 2-3 years and cannot produce immune memory, resulting in the failure of vaccine development. So far, only 6 potential coronavirus vaccines have entered the clinical research stage, but no effective vaccine has been approved for marketing

Method used

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  • Nano coronavirus recombinant vaccine taking graphene oxide as carrier
  • Nano coronavirus recombinant vaccine taking graphene oxide as carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Preparation process of graphene oxide (GO)-carnosine-CpG-RBD recombinant protein vaccine preparation

[0045] The TLR9 receptor nucleic acid sequence CpG ODN M362, which has cross-reactivity to both humans and mice, is selected. The specific sequence is as follows: 5'-TCGTCGTCGTTC:GAACGACGTTGAT-3' (25 mer, SEQ ID NO 1), using the improved method of EDC-NHS reaction For the coupling process of GO and carnosine, 26 mg of GO freeze-dried powder was added to 5.20 mL of phosphate buffered saline (PBS, pH = 7.4), and sonicated (200 W, 40 kHz) at 25°C for 3 h . Add 6.82 mg EDC (N1-((ethylimino)methylene)-N3,N3-dimethylpropane-1,3-diamine at 25°C, Chinese: 1-(3-dimethylaminopropyl)-3-ethylcarbon diimide) and 7.73 mg NHS (N-N-hydroxysuccinimide) to activate the GO solution. Excess EDC / sulfo-NHS was removed from the reaction solution by ultrafiltration, and then the pH of the solution was adjusted to 7.4. Then, 40 mg of carnosine, 1.2ug of CpG, and different concentrations of ...

Embodiment 2

[0047] Graphene oxide (GO)-carnosine-CpG-RBD recombinant protein vaccine immunization experiments in mice

[0048] according to figure 1 The marked schedules were immunized with 6-week-old female BALB / cmice mice by subcutaneous injection on days 0, 14, and 28, until day 28 and day 42. Blood was collected by subcutaneous injection, serum was separated, and serum was detected. Specific antibodies against RBD. Mice were sacrificed on day 42, splenocytes were isolated, and the specific T cell immune response and cytokine secretion were detected.

[0049] Grouping and dose determination of immunized mice:

[0050] 1. (Graphene oxide+carnosine)+ 1.2ug cpG+3ug RBD

[0051] 2. (Graphene oxide+carnosine)+ 1.2ug cpG+6ug RBD

[0052] 3. Aluminum hydroxide + 6ug RBD (1:1)

[0053] 4. 6ug RBD

[0054] 5. Liposome (lipo) + 6ug RBD group

[0055] Mouse strain: BALB / c mice (n=6).

[0056] The schedule for immunizing mice with GO-Car-carnosine-CpG-RBD vaccine is: blood collection and f...

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Abstract

The invention belongs to the field of nano materials and biological medicines, and relates to a vaccine, in particular to development of a 2019-nCoV coronavirus nuclear recombinant nano vaccine. The invention also comprises a preparation method of the vaccine and application of the vaccine in animal experiments. The novel coronavirus vaccine contains graphene oxide, carnosine, CpG and novel coronavirus RBD; The carnosine, the CpG and novel coronavirus RBD are combined on a framework of the graphene oxide; the coding sequence of the CpG is as shown in SEQ ID NO 1; and the novel coronavirus RBDrefers to that a novel coronavirus protein receptor binding region can generate a high-titer specific antibody aiming at the RBD in a mouse body, and strong support is provided for prevention and treatment of the novel coronavirus.

Description

technical field [0001] The invention belongs to the field of nanomaterials and biomedicine, and relates to the development of a vaccine development platform. Specifically, it involves the development of nuclear recombinant nano-vaccine for 2019-nCoV coronavirus. The present invention also includes the application of the vaccine in animal experiments. [0002] technical background [0003] Vaccines are the ultimate weapon to eradicate major infectious diseases. Compared with other treatments, vaccines have the lowest cost and have the advantage of being the first to strike the enemy. They have undoubtedly become the hope of the public. Through vaccination, human beings have eliminated smallpox and polio cases The incidence of infectious diseases such as diphtheria is rare, and the incidence of diseases such as measles and neonatal tetanus has dropped significantly. The impact of vaccines on human health cannot be overstated. The birth of each new vaccine is a great victory f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61K39/39A61K39/385A61P31/14C07K14/165C07K17/14
CPCA61K39/12A61K39/39A61K39/385A61P31/14C07K14/005C07K17/14C12N2770/20034A61K2039/55561A61K2039/627A61K2039/60C12N2770/20022
Inventor 崔大祥高昂梁辉田静李雪玲沈琦
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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