CQD (Carbon Quantum Dot) adjuvant and vaccine containing same

A technology for carbon quantum dots and vaccines, applied in vaccines, veterinary vaccines, medical preparations containing active ingredients, etc., can solve problems such as unfavorable injection, limited combination methods, poor emulsification, etc. Long-lasting immune memory and effective immune protection

Inactive Publication Date: 2018-09-25
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology uses certain types of molecules called quantum dots (also known as semiconductor nanocrystals) or other substances like zinc oxide particles to create small objects which when exposed to light activate specific receptors on nearby cells they release their energy into surrounding water causing them to emit green fluorescences over several hours later. These tiny red lights indicate how well these special chemical compounds were able to stimulate humoral defense mechanisms such as BCGA from bacteria infected fish eggs. By measuring this reaction at different locations around the seafood samples we could determine whether there was any harm caused due to ingestion of contaminated food containing harmful microorganism(such as Escherichia coli).

Problems solved by technology

The technical problem addressed in this patented text relates how safe and efficient administration of certain types of drugs requires developing novel ways to improve their effectiveness without causing negative reactions like inflammatory ones due to lack of sufficient immunity from these agents against targeted molecules.

Method used

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  • CQD (Carbon Quantum Dot) adjuvant and vaccine containing same
  • CQD (Carbon Quantum Dot) adjuvant and vaccine containing same
  • CQD (Carbon Quantum Dot) adjuvant and vaccine containing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Preparation of carbon quantum dots

[0029] Humic acid (0.2 g) was dissolved in 50 mL of secondary water and transferred to an autoclave with Teflon. The autoclave was heated at 180°C for 5 hours and cooled naturally to obtain a dark brown solution. The reaction solution was centrifuged at 3000 rpm for 15 minutes. Subsequently, the precipitate was discarded, and the supernatant was dialyzed in a dialysis bag (MWCO=1000) for 48 hours to remove small molecule impurities. Afterwards, the supernatant was centrifuged at 12000 rpm for 15 minutes, and the obtained fluorescent CQDs were dried in a vacuum oven and stored in a refrigerator at 4 °C.

[0030] The size and morphology of the synthesized CQDs were studied by transmission electron microscopy (TEM), as figure 1 shown. Depend on figure 1 It can be seen that the CQDs particles prepared in this example are spherical with a diameter of 3-5 nm.

Embodiment 2

[0031] Embodiment 2: Expression and identification of ALV-J gp85 protein

[0032] Firstly, the recombinant plasmid (PET28a-gp85) was transformed into Rosetta (DE3) cells, and then induced with 0.5mM IPTG for 6 hours at 37°C to obtain the recombinant gp85 protein. The recombinant gp85 protein was purified by high-affinity Ni-NTA column and identified by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) after dialysis ( figure 2 a). Proteins were subjected to gp85-specific M antibody JE9 under Western blot analysis ( figure 2 b). The concentration of gp85 protein collected by PEG6000 was determined to be 1.2 mg / mL by thin layer chromatography.

Embodiment 3

[0033] Example 3: Effect of carbon quantum dots on the activity of chicken lymphocytes

[0034] MTT assay was used to determine the effect of CQDs on the viability of chicken lymphocytes. details as follows:

[0035] Chicken lymphocytes were grown in 96-well microplates in Roswell Park Memorial Institute medium containing 10% fetal bovine serum and 1% penicillin / streptomycin in 5% CO 2 Incubate at 37°C for 24 hours. Solutions of synthetic CQDs (prepared in Example 1) at different concentrations were loaded into each well; each concentration was prepared in triplicate and incubated for 24 hours. Afterwards, 20 mL of 5 mg / mL 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) solution was added to each well containing cells and incubated Incubate for 4 hours. The medium was removed and 150 mL of DMSO was added to release the methyl groups. The mixture was shaken for about 15 minutes, and the luminosity (OD) was measured by a microplate reader. Cell viability w...

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Abstract

The invention discloses a CQD (Carbon Quantum Dot) adjuvant and a vaccine containing the same, i.e., each single part of vaccine agent contains 200 to 400mug of the CQD adjuvant. The phenomenon that CQDs can be used as an excellent adjuvant for resisting ALV-J (avian leukosis virus subgroup J) is found for the first time; researches find that the CQDs prepared by the invention have no cytotoxicity, cannot induce reduction of the cell activity, are good in biocompatibility, and are safe and reliable to use in the immunizing dose range. The vaccine prepared by using the CQDs as the adjuvant, which is disclosed by the invention, with respect to a vaccine prepared by a Freund's adjuvant, can more effectively induce the generation of a high antibody titer to carry out immune protection and provide a more lasting protective immune response.

Description

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Claims

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Application Information

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Owner SHANDONG AGRICULTURAL UNIVERSITY
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