Supercharge Your Innovation With Domain-Expert AI Agents!

Application of trichosanthin in sensitization and/or activation of dendritic cells

A technology of dendritic cells and trichosanthin, applied in the field of biomedicine, can solve problems that are difficult to prevent the occurrence and development of tumors

Pending Publication Date: 2019-05-28
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are a series of immune surveillance mechanisms in the body, it is still difficult to prevent the occurrence and development of tumors

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 trichosanthin in sensitization and/or activation of dendritic cells
  • Application of trichosanthin in sensitization and/or activation of dendritic cells
  • Application of trichosanthin in sensitization and/or activation of dendritic cells

Examples

Experimental program
Comparison scheme
Effect test

preparation Embodiment 1

[0204] Preparation of fusion protein 1 (TCS-antigen peptide protein)

[0205] Prokaryotic expression and purification of TCS-antigen peptide protein

[0206] a: The constructed plasmid TCS-antigen peptide was transformed into Escherichia coli BL21(DE3) competent cells.

[0207]b: Transfer the strain containing the recombinant plasmid TCS-antigen peptide to LB medium containing 100 μg / ml Amp, and culture it in a constant temperature shaker at 37°C at 220rpm until the logarithmic growth phase (absorbance value at 600nm is 0.6-0.8), add IPTG with a final concentration of 1 mM was expressed overnight (16 h) at 25° C. and 150 rpm.

[0208] c: Use a centrifuge at 4°C and 9,000 rpm for 3 minutes to collect bacteria.

[0209] d: Resuspend the bacteria in HEPES buffer (containing 20 mM HEPES, 150 mM NaCl, 1 mM EDTA, 0.5‰ Tween 20, pH 8.5).

[0210] e: Use a probe sonicator with a power of 400 W to sonicate the cells for 35 minutes.

[0211] f: 12,000rpm, centrifuge at 4°C for 30min...

preparation Embodiment 2

[0217] Preparation of fusion protein 2 (penetrating peptide-TCS-antigen peptide protein)

[0218] Prokaryotic expression and purification of penetrating peptide-TCS-antigen peptide protein

[0219] a: The constructed plasmid-penetrating peptide-TCS-antigen peptide was transformed into Escherichia coli BL21(DE3) competent cells.

[0220] b: The strain containing the recombinant plasmid penetrating peptide-TCS-antigen peptide was transferred to the LB medium containing 100 μg / ml Amp, and cultured in a constant temperature shaker at 37°C at 220rpm to the logarithmic growth phase (absorbance value at 600nm was 0.6- 0.8), adding IPTG with a final concentration of 1 mM, and expressed overnight (16 h) at 37° C. and 220 rpm.

[0221] c: Use a pre-cooled centrifuge at 4°C and 9,000 rpm for 3 minutes to collect the bacteria.

[0222] d: Resuspend the bacteria in HEPES buffer (containing 20 mM HEPES, 150 mM NaCl, 1 mM EDTA, 0.5‰ Tween 20, pH 8.5).

[0223] e: The cells were sonicated ...

experiment Embodiment 1

[0230] MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyl bromotetrazolium blue, trade name: thiazolium blue) method for the determination of recombinant protein drugs TCS, TCS-antigenic peptide, Cytotoxic effect of penetrating peptide-TCS-antigen peptide Digest and count murine dendritic cells DC2.4 cells in the logarithmic growth phase, and dilute to a density of 4×10 4 The cell suspension of 1 cell / mL was transferred to a 96 cell culture well plate, and 100 μL of cell suspension was added to each well, and cultured with DMEM complete medium containing 10% calf serum for 12 h (37 °C, 5% CO 2 ).

[0231] Determine the optimal drug concentration range through preliminary experiments, add solutions with different concentrations, and make 6 replicate holes for each concentration. After culturing for 48 hours, 20 μl of MTT (5 mg / ml, purchased from Sigma-Aldrich, USA) was added and incubated for 4 hours. The culture supernatant was carefully aspirated, 200 μL of DMSO was added to each ...

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

No PUM Login to View More

Abstract

The invention provides application of trichosanthin in sensitization and / or activation of dendritic cells. In particular, the invention provides uses of trichosanthin (TCS) or a coding gene thereof for preparing a dendritic cell activator and / or sensitizer. The trichosanthin can effectively activate and / or sensitize the dendritic cells, thereby enhancing the antigen-presenting ability thereof; after a fusion protein containing a tumor antigen and trichosanthin can be processed and presented by the dendritic cells, the immune system against tumors in vivo can be specifically activated, and thetumor growth and diffusion are inhibited, thereby the fusion protein becomes a therapeutic vaccine for tumors. The trichosanthin is highly safe and has small toxic and side effects, and is an ideal component of a tumor therapeutic vaccine preparation.

Description

technical field [0001] The invention belongs to the field of biomedicine, and more specifically, the invention relates to the application of trichosanthin in activating dendritic cells, and a fusion protein containing trichosanthin and tumor antigen prepared by utilizing the characteristic. Background technique [0002] Malignant tumors seriously threaten human health. With the change of people's living habits, the morbidity and mortality of tumors are on the rise. Tumor treatments usually include surgical resection, chemotherapy, radiotherapy, and immunotherapy. [0003] Although great progress has been made in cancer drug therapy in recent years, the application of macromolecular drugs in cancer treatment is less. With the development of biotechnology, macromolecular drugs, especially protein drugs, have received more and more attention. [0004] Tumor is the product of malignant transformation of normal cells in the body, which is characterized by continuous proliferat...

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
IPC IPC(8): C12N5/0784C07K19/00C12N15/62C12N15/70C12N1/21A61K39/00A61P35/00
CPCA61K38/16A61K39/00A61K47/64A61P35/00C07K14/415C07K19/00C12N15/62C12N15/70A61K39/4644A61K39/4615A61K39/4622A61K2239/50
Inventor 黄永焯吴爱花陈应之
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com