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AIDS live carrier multipartial vaccine and construction method thereof

A multivalent vaccine and AIDS technology, applied in the field of AIDS vaccine, can solve the problems of insufficient immunogenicity and low antigen expression, and achieve the effects of low cost, strong antigenicity, and simple preparation and purification methods

Inactive Publication Date: 2010-06-02
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes an improved method that allows small molecules called silver nanospikes (AgNPs) attached on tiny fibers made from synthetic resins like polyacrylamide gel particles. These minute nano spots contain many different types of DNA segments connected by short peptides. When these DNA segments come together they create large structures containing thousands of smaller pieces of DNA. Silver Nspores were found effective at killing certain germ cells but also affect other organisms' ability to fight off harmful microorganism such as viruses. They could potentially use this technique to develop new ways to prevent diseases caused by virus spread through infected individuals.

Problems solved by technology

The technical problem addressed in this patents relating to developing safe and effective methods for producing specific types of peptides called neutralized epimicrobial antigens (NepiV), particularly those specifically associated with helper functions like cytidylic acid transcripts (CLTs)/cytotoxins (CTLs)-related ones. This approach could potentially lead to improved therapies for treatments related to AIDs.

Method used

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  • AIDS live carrier multipartial vaccine and construction method thereof
  • AIDS live carrier multipartial vaccine and construction method thereof
  • AIDS live carrier multipartial vaccine and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0045] Example 2 Integration of 2F5 epitope sequence into S.typhimurium ST14028-3b genome

[0046] Gene replacement: The constructed plasmid PHSG2F5A6 was transformed into S. typhimurium ST14028-3b by electroporation. Pick a single colony that has been confirmed to contain the recombinant plasmid PHSG2F5A6 and inoculate it into Terrific broth containing ampicillin 100ug / ml for overnight culture at 42°C. Dilute the culture appropriately and spread it on an LB plate containing ampicillin 100ug / ml at 42°C Cultivate for 18 hours; pick 4-8 ​​confirmed ampicillin-resistant colonies, mix and inoculate them into 5ml Terrific broth, and culture overnight at 28°C; take 5ul of the overnight culture broth and inoculate into a new 5ml Terrific Cultivate overnight at 28°C in broth, repeat this step 3 times; spread on LB agar plate after diluting about 10-6 to 10-8, and cultivate overnight at 28°C; On LB agar plates and LB agar plates without antibiotics, cultivate overnight at 28°C, and se...

Embodiment 3

[0049] Example 3 identifies the expression of the 2F5 epitope.

[0050] Western blotting analysis: Pick a single colony of the SA-2F5 strain containing the exogenous fragment 2F5 after the gene replacement is completed, culture it overnight at 37°C in 1ml LB liquid medium, centrifuge at 12,000 rpm for 1 minute, remove the supernatant, and resuspend in In 200ul SDS sample buffer, boil for 10min, centrifuge at 13200 rpm for 10 minutes, wash the precipitate with 500ul deionized water three times, add 100ul formic acid to resuspend, freeze in a -80°C ultra-low temperature refrigerator, vacuum dry, add 15ul SDS sample buffer Resuspended, boiled for 10 min, and loaded for SDS-PAGE electrophoresis. SDS-PAGE electrophoresis uses 5% stacking gel and 15% separating gel. After electrophoresis, the separated proteins were transferred to PVDF membranes, and Western blotting was performed using antibacterial hair SEF17 rabbit serum and anti-rabbit horseradish peroxidase-labeled IgG seconda...

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Abstract

The invention discloses an AIDS live carrier multipartial vaccine and a construction method thereof. The AIDS live carrier multipartial vaccine takes fine polymerized pilus of S. typhimurium as a live carrier of the vaccine, so that the epitope of AIDS virus can be expressed on the fine polymerized pilus of S. typhimurium. The invention takes the fine polymerized pilus of S. typhimurium as the live carrier of the vaccine, and the constructed live carrier multipartial vaccine has high antigen express and good immunogenicity and can effectively prevent AIDS virus, wherein the high antigen express means that neutralizing antibodies and specific T cells which have enough titers can be induced.

Description

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Claims

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Application Information

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Owner HARBIN MEDICAL UNIVERSITY
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