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

Application of Streptococcus pneumoniae protein in resisting Streptococcus pneumoniae infection

A streptococcus pneumoniae and protein technology, applied in the direction of immunoglobulin, antibacterial immunoglobulin, application, etc., can solve the problems of adverse reactions and insufficient immunogenicity, so as to improve the body's IgG antibody response and enhance resistance to pneumonia chain Effects of bacterial infection

Active Publication Date: 2022-02-22
CHONGQING MEDICAL UNIVERSITY
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the above-mentioned shortcoming of the prior art, the object of the present invention is to provide the application of Streptococcus pneumoniae protein in the anti-Streptococcus pneumoniae infection, be used to solve its immunogenicity of most proteins in the prior art still not strong enough, can cause Adverse reactions and other issues

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
  • Application of Streptococcus pneumoniae protein in resisting Streptococcus pneumoniae infection
  • Application of Streptococcus pneumoniae protein in resisting Streptococcus pneumoniae infection
  • Application of Streptococcus pneumoniae protein in resisting Streptococcus pneumoniae infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Recombinant expression plasmid pET28a(+)-PepO, pET28a(+)-ZmpB 673-863 , pET28a(+)-ZmpB 673-863 -PepO and pET28a(+)-PepO-ZmpB 673-863 Construction of expression vectors.

[0089] (1) Materials:

[0090] Plasmid pET28a(+) was purchased from Novagen, Prime Star high-fidelity enzyme, dNTPs, Buffer, MgCl used in PCR 2 Purchased from Bao Biological Engineering (Dalian) Co., Ltd., PTC-200 PCR instrument was a Perkin Elmer product, and fluorescent quantitative PCR instrument RG-3000 was purchased from Corbett Research.

[0091] (2) Design and synthesis of primers:

[0092] Using Streptococcus pneumoniae D39 genomic DNA as a template, refer to its complete sequence (GeneBank number CP000410.2), use Premier5.0 to design primers, and synthesize them by Sangon Bioengineering (Shanghai) Co., Ltd.

[0093] ZmB 673-863 : Upstream primer: 5'-GCCATGGTTGAAGAAGTTGTTGTT-3', containing NcoI site;

[0094] Downstream primer: 5'-CCCTCGAGATCTCCAAGACTGTTAAT-3', containing XhoI site;

[...

Embodiment 2

[0162] Prokaryotic expression plasmids pET28a(+)-PepO, pET28a(+)-ZmpB 673-863 , pET28a(+)-ZmpB 673-863 -PepO and pET28a(+)-PepO-ZmpB 673-863 Expression, identification and purification in Escherichia coli.

[0163] (1) Recombinant plasmids pET28a(+)-PepO, pET28a(+)-ZmpB 673-863 , pET28a(+)-ZmpB 673-863 -PepO and pET28a(+)-PepO-ZmpB 673-863 Transformed into the host strain BL21(DE3).

[0164] (2) IPTG induces PepO and ZmpB 673-863 , ZmpB 673-863 -PepO (fusion protein) and PepO-ZmpB 673-863 (Fusion protein) expression in large quantities.

[0165] (3) Purification of the recombinant protein: After the bacteria were broken by ultrasonic, the supernatant of the broken bacteria was taken for purification; 4°C, 10000rpm×10min, the supernatant was filtered with a 0.45μm membrane filter, and the filtrate was collected for later use.

[0166] Affinity chromatography purification: pipette 2ml of 50% Ni 2+ -NTA resin suspension in the chromatography column, equilibrate with 20m...

Embodiment 3

[0180] Detection of ZmpB by Western blot 673-863 Recognition reaction of protein antiserum to Streptococcus pneumoniae bacterial protein.

[0181] (1) Streptococcus pneumoniae CMCC 31109 (type 1), D39 (type 2), CMCC 31436 (type 3), TIGR4 (type 4), CMCC 31207 (type 6B), CMCC31507 (type 7F), CMCC31216 (type 9V) , CMCC 31614 (Type 14), CMCC31687 (Type 18C), CMCC31693 (Type 19F) and CMCC31759 (Type 23F). A total of 11 different serotypes of bacteria were inoculated in 30ml C+Y medium, 37°C, 5% CO 2 Cultivate until the bacteria are in the logarithmic growth phase OD600=0.4-0.6, collect the bacteria, centrifuge at 12000g for 5min, wash twice with sterile PBS, resuspend the bacterial cell pellet with 100μl PBS, add 5×loadingbuffer in proportion, boil in boiling water for 30min, Centrifuge at 12000g for 5min, and take the supernatant for Western Blot analysis.

[0182](2) PAGE electrophoresis: the loading amount of protein per hole is 2 μg, 8% separating gel+5% stacking gel, 80V, 3...

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
Login to View More

Abstract

The invention provides the application of Streptococcus pneumoniae protein in resisting Streptococcus pneumoniae infection. The Streptococcus pneumoniae endopeptidase O (PepO) of the present invention is a subcutaneous immune adjuvant, mixed and fused with the 673rd to 863rd amino acid peptide of zinc metalloprotease B (ZmpB), and the prepared Streptococcus pneumoniae Protein vaccines have a good protective effect against Streptococcus pneumoniae infection.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to Streptococcus pneumoniae PepO, ZmpB 673-863 Use in the fight against Streptococcus pneumoniae infection, especially related to the use of Streptococcus pneumoniae endopeptidase O as an adjuvant for subcutaneous immunization, ZmpB 673-863 Use as a vaccine alone in combination with other adjuvants and PepO and ZmpB 673-863 Application of mixed or fusion expression in anti-streptococcus pneumoniae infection. Background technique [0002] Streptococcus pneumoniae can cause pneumonia, meningitis and sepsis, and nearly 400,000 children under the age of 5 die every year worldwide due to Streptococcus pneumoniae infection. Pneumococcal sepsis is the leading cause of infant mortality in developing countries, where approximately 25% of deaths of children under 5 years of age and more than 1.2 million infant deaths are preventable each year. The current conjugate vaccine formulations...

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 Patents(China)
IPC IPC(8): C07K14/315C12N15/31C12N15/70C12N1/21C07K16/12A61K39/09A61P31/04
CPCA61K39/092A61P31/04C12N15/70C07K14/3156C07K16/1275A61K38/00
Inventor 尹一兵尹一帆张雪梅胥文春王虹何於娟肖江明王婧瑶
Owner CHONGQING MEDICAL UNIVERSITY
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