Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Subunit vaccine against Mycobacterium tuberculosis based on modification by arabinogalactan-polyinosinic acid polycytidylic acid and preparation method thereof

A technology of polyinosinic acid cytidylic acid and arabinogalactan, which is applied in the field of Mycobacterium tuberculosis subunit vaccines, can solve problems such as toxic and side effects, and achieve the effects of preventing tuberculosis, enhancing immune response, and prolonging circulation

Inactive Publication Date: 2016-08-31
INST OF PROCESS ENG CHINESE ACAD OF SCI
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high doses of poly(I:C) may induce autoimmune reactions and produce some other toxic side effects, so the dose of poly(I:C) needs to be strictly controlled in clinical applications

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Subunit vaccine against Mycobacterium tuberculosis based on modification by arabinogalactan-polyinosinic acid polycytidylic acid and preparation method thereof
  • Subunit vaccine against Mycobacterium tuberculosis based on modification by arabinogalactan-polyinosinic acid polycytidylic acid and preparation method thereof
  • Subunit vaccine against Mycobacterium tuberculosis based on modification by arabinogalactan-polyinosinic acid polycytidylic acid and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Preparation of CT fusion protein

[0024] (1) Construction of Ag85B-HspX recombinant plasmid

[0025] First, according to the Ag85B coding gene sequence of the standard Mycobacterium tuberculosis strain Rv1886c and the HspX coding gene sequence of Rv2031c in GenBank, three Gly-Gly-Gly-Gly-Gly-Ser (Gly-Gly-Gly-Ser) repeats were designed in the middle The whole gene sequence of the unit was then synthesized; then primers were designed, the whole gene sequence was amplified by PCR, the target gene was recovered with a gel recovery kit, and the whole gene sequence was double-digested with HindⅢ and MscⅠ. At the same time, the carrier plasmid pET26b was double-enzymatically digested, and after the target gene fragments were recovered separately, they were ligated with T4 ligase; finally, the ligated products were transformed into E. Coli BL21 (DE3), positive clones were screened by colony PCR and sequenced for identification, thereby The pET26b-Ag85B-HspX reco...

Embodiment 2

[0034] Embodiment 2: Preparation of Mycobacterium tuberculosis subunit vaccine

[0035] (1) Derivation of poly(I:C)

[0036] Weigh 9.98g of ethylenediamine and 2.60g of sodium bisulfite, dissolve in 25ml of deionized water, mix well, then adjust the pH of the solution to 5.5-6.0 with sodium hydroxide, add 12.5mg of poly(I:C), well mixed. Then react at 42°C for 3 hours ( figure 2 ). After the reaction, the reaction solution was fully dialyzed in deionized water with a dialysis bag with a molecular weight cut-off of 8-12 kDa, and then freeze-dried for future use.

[0037] (2) Preparation of AG-P conjugates

[0038] Weigh 10 mg AG and dissolve in 20 mM acetic acid-sodium acetate buffer solution (pH 5.5-6.0). Sodium periodate was added to make the final concentration 20 mM, and after reaction at room temperature in the dark for 20 minutes, a small amount of ethylene glycol was added to terminate the reaction. Subsequently, it was dialyzed three times for 12 hours in 20 mM p...

Embodiment 3

[0045] Example 3: Structural Characterization of Vaccines

[0046] The three vaccines were identified by analytical Superdex 200 gel filtration column (1.0×30cm), the eluent was 20mM phosphate buffer (pH 7.4) containing 0.15M sodium chloride, and the flow rate was 0.5ml / min. Such as image 3 As shown, compared with the fusion protein AH, the peak times of AH-AG, AH-P and AH-AG-P were all significantly earlier. Among them, the peak time of AH-AG-P was earlier than that of AH-AG and AH-P, indicating that the modification of AH by AG-P conjugates significantly increased the molecular weight of AH.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention provides a subunit vaccine against Mycobacterium tuberculosis based on modification by arabinogalactan-polyinosinic acid polycytidylic acid and a preparation method thereof. The preparation method comprises the following steps: subjecting arabinogalactan (AG) and polyinosinic acid polycytidylic acid (poly(I:C)) to chemical coupling so as to prepare an AG-poly(I:C) conjugate which is used as an immunoadjuvant; subjecting antigen protein Ag85B expressed in a replicative period of Mycobacterium tuberculosis and antigen protein HspX expressed in an incubation period of Mycobacterium tuberculosis to recombinant fusion expression in Escherichia coli so as to obtain Ag85B-HspX fusion protein; and modifying the Ag85B-HspX fusion protein with the AG-poly(I:C) conjugate so as to obtain the subunit vaccine against Mycobacterium tuberculosis. The vaccine has a long cycle half-life period in a body and can stimulate the body to generate high-level humoral immune response and cellullar immune response.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a mycobacterium tuberculosis subunit vaccine based on arabinogalactan-polyinosinic acid cytidylic acid modification. Background technique [0002] Tuberculosis (Tuberculosis) is a very old infectious disease caused by Mycobacterium tuberculosis, and it is also one of the main death threats for HIV-infected people. According to the World Health Organization, about one-third of the world's population is infected with Mycobacterium tuberculosis, and there are about 8.7 million new tuberculosis cases and 1.5 million tuberculosis deaths every year. Due to the long-term abuse of anti-tuberculosis drugs (isoniazid, streptomycin, rifampicin and ethambutol, etc.), the cases of infection with multidrug-resistant and extensively drug-resistant strains have increased year by year. Therefore, the problem of drug resistance of Mycobacterium tuberculosis has become very serious. my country is the s...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/04A61K39/39A61P31/06
CPCA61K39/04A61K39/39A61K2039/55561A61K2039/575
Inventor 胡涛于卫立黄庆瑞
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com