Method for improving thermal stability of virus-like particles based on metal organic framework biomimetic mineralization

A metal organic framework and thermal stability technology, applied in the field of medicine, can solve the problems of refrigerated transportation, poor thermal stability, etc., and achieve the effects of enhanced sedimentation coefficient, increased encapsulation capacity, and enhanced cellular immune response level.

Pending Publication Date: 2022-05-06
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The purpose of the present invention is to provide a method for improving the thermal stability of virus-like particle vaccines based on metal-organic framework biomimetic mineralization, which is used to solve the problem of poor thermal stability of existing VLPs vaccines and the need for refrigerated transportation

Method used

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  • Method for improving thermal stability of virus-like particles based on metal organic framework biomimetic mineralization
  • Method for improving thermal stability of virus-like particles based on metal organic framework biomimetic mineralization
  • Method for improving thermal stability of virus-like particles based on metal organic framework biomimetic mineralization

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Effect test

Embodiment 1

[0044] The synthesis of embodiment 1FMDV-VLPs-ZIF-8

[0045] (1) To 1 part by volume, the content of virus-like particles is 400-600 μg / mL foot-and-mouth disease virus-like particles (FMDV-VLPs) solution to be encapsulated (the virus-like particles are 201610929279.0 according to the application number, and the invention name is "one 4.5 parts by volume of 80mM dimethylimidazole solution were added to the O-type foot-and-mouth disease virus-like particle and its preparation method and application "patent application" and fully mixed;

[0046] (2) Add 0.5 parts by volume of 20mM zinc nitrate solution to the mixed solution of step (1), mix well, and react for 10min;

[0047] (3) After the reaction in step (2) is complete, add 4.5 parts by volume of 80mM dimethylimidazole solution and 0.5 parts by volume of 20mM zinc nitrate solution to mix and react for 20min;

[0048] (4) After the reaction in step (3) is complete, after ultrasonic dispersion, add 0.1-0.2 parts by volume of 2M...

Embodiment 2

[0052] Identification of embodiment 2FMDV-VLPs-ZIF-8 complex

[0053] 1. Identification of complexes by Fourier infrared spectroscopy

[0054] After freeze-drying the synthesized ZIF-8, FMDV-VLPs-ZIF-8 and protein, add an appropriate amount of fully dried, use Fourier infrared spectrometer to measure the absorption peak, and determine the encapsulation situation through the absorption peak. The result is as figure 2 As shown in A, the results show that the FMDV-VLPs-ZIF-8 complex has specific absorption peaks of ZIF-8 and protein, indicating that FMDV-VLPs are successfully encapsulated.

[0055] 2. Identification of complexes by SDS-PAGE

[0056] The FMDV-VLPs before encapsulation and the mineralized encapsulation FMDV-VLPs-ZIF-8 samples were subjected to denaturing electrophoresis, and the protein gel was stained with Coomassie brilliant blue staining solution, and decolorized to observe the encapsulation of FMDV-VLPs by metal organic framework mineralization Happening. ...

Embodiment 3

[0061] Example 3 Heat resistance analysis of FMDV-VLPs-ZIF-8

[0062] To study the thermal stability of FMDV-VLPs and FMDV-VLPs-ZIF-8 composites, 1 mL samples were treated at 45°C (1h, 3h, and 6h) and 60°C (0.5h, 1h, and 3h), and untreated FMDV-VLPs and FMDV-VLPs-ZIF-8 composites served as controls. Samples were collected at predetermined times and placed on ice. After the samples were hydrolyzed with EDTA solution overnight, the protein released into the supernatant was collected by centrifugation (8000 rpm, 10 min), and the protein content in the supernatant was measured using the Micro BCA assay. . The protein activity of FMDV-VLPs was determined by ELISA. The result is as image 3 As shown, the results show that metal-organic framework encapsulation of FMDV-VLPs can improve the heat resistance of FMDV-VLPs.

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Abstract

The invention discloses a method for improving thermal stability of virus-like particles (VLPs) based on metal organic framework biomimetic mineralization. According to the preparation method disclosed by the invention, VLPs is successfully encapsulated in a zeolite imidazole framework 8 (ZIF-8) under the coordination action of dimethylimidazole and zinc nitrate, so that the VLPs-ZIF-8 nanoparticles are formed. According to the invention, a frame structure formed by coordination of metal ions and organic matters is encapsulated outside the virus-like particles, so that the heat resistance of the VLPs vaccine is improved, and the problems of consumption of a large amount of manpower and material resources caused by cold chain transportation and the like are solved. The prepared ZIF-8 metal organic framework is low in cost, the method is simple, and the synthesis process is mild. The cellular immune response level of the VLPs vaccine biomimetically mineralized by the metal organic framework is obviously improved, and a new technical means is provided for improving the cellular immune level of the VLPs vaccine.

Description

technical field [0001] The invention relates to a method for improving the thermal stability of virus-like particles, in particular to a method for improving the thermal stability of virus-like particles based on metal-organic framework biomimetic mineralization. The invention belongs to the technical field of medicine. Background technique [0002] In recent years, virus like particles (VLPs) have played a vital role in the fields of disease prevention, control, immunotherapy and nanotechnology research. VLPs are composed of the main structural proteins of the virion. They do not contain the virus genome and have high safety. Moreover, VLPs have a structure similar to natural viruses and are easily recognized and taken up by antigen-presenting cells (APCs), thereby inducing a good immune response. Excellent immunogenicity and reactogenicity. Therefore, VLPs have become a hotspot in modern vaccine research because of their own safety, ease of production, and immune charact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09C07K17/08C07K17/04A61K39/135A61K39/385A61P31/14
CPCC07K14/005C07K17/08C07K17/04A61K39/12A61K39/385A61P31/14C12N2770/32123C12N2770/32134C12N2770/32151A61K2039/5258A61K2039/552
Inventor 郭慧琛孙世琪滕志东白满元张韵宋河涛李家俊董虎吴金恩丁耀忠尹双辉
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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