A new der p1 nano vaccine for the treatment of lung cancer and its preparation method and use

A nano-vaccine and lung cancer technology, applied in the direction of medical preparations containing active ingredients, pharmaceutical formulas, antibody medical components, etc., to achieve the effect of increasing the body's immunity, good anti-cancer function, and significant therapeutic effect

Active Publication Date: 2018-04-10
NANHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention has not been reported both at home and abroad

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A new Der p1 nano-vaccine for treating lung cancer is prepared by adopting the following steps:

[0035] Step 1, using genetic engineering technology to prepare the recombinant house dust mite allergen Der p1:

[0036] (1) Pick a single genetically engineered bacterial colony containing the recombinant expression plasmid recombinant pET-24a(+)Der p1 and inoculate it in 20 ml of LB containing 50 μg / ml kanamycin, shake the bacteria overnight at 35°C, 160 rpm, and press 1% Inoculate the genetically engineered bacterial colony in 1L LB / kam culture solution, shake the bacteria at 200rpm until the OD600 is 0.5, add IPTG to make the final concentration 0.1mM, after induction for 4 hours, centrifuge at 2000g for 5min at 2 °C, remove The supernatant, the pellet was washed once with 1xPBS, the bacteria were resuspended in 40ml 1xPBS / 5mM EDTA, lysomycetes were added to a final concentration of 0.4mg / ml, DTT to a final concentration of 5mM, PMSF to a final concentration of 0.8mmol / ...

Embodiment 2

[0043] A new Der p1 nano-vaccine for treating lung cancer is prepared by adopting the following steps:

[0044] Step 1, using genetic engineering technology to prepare the recombinant house dust mite allergen Der p1:

[0045] (1) Pick a single genetically engineered bacterial colony containing the recombinant expression plasmid recombinant pET-24a(+)Der p1 and inoculate it in 20 ml of LB containing 50 μg / ml kanamycin, shake the bacteria overnight at 37°C, 240 rpm, and press 4% Inoculate the genetically engineered bacterial colony in 1L LB / kam culture solution, shake the bacteria at 300 rpm until the OD600 is 0.7, add IPTG to make the final concentration 0.4 mM, after induction for 6 hours, centrifuge at 4000 g at 6 °C for 15 min, remove The supernatant, the pellet was washed once with 1xPBS, the bacteria were resuspended in 40ml 1xPBS / 5mM EDTA, lysomycetes were added to a final concentration of 0.6mg / ml, DTT to a final concentration of 15mM, PMSF to a final concentration of 1....

Embodiment 3

[0052] A new preparation method of Der p1 nano-vaccine for treating lung cancer, the steps are as follows:

[0053] Step 1, using genetic engineering technology to prepare the recombinant house dust mite allergen Der p1:

[0054] (1) Pick a single genetically engineered bacterial colony containing the recombinant expression plasmid recombinant pET-24a(+)Der p1 and inoculate it in 20 ml of LB containing 50 μg / ml kanamycin, shake the bacteria overnight at 35°C, 160 rpm, and press 1% Inoculate the genetically engineered bacterial colony in 1L LB / kam culture solution, shake the bacteria at 200rpm until the OD600 is 0.5, add IPTG to make the final concentration 0.1mM, after induction for 4 hours, centrifuge at 2000g for 5min at 2 °C, remove The supernatant, the pellet was washed once with 1xPBS, the bacteria were resuspended in 40ml 1xPBS / 5mM EDTA, lysomycetes were added to a final concentration of 0.4mg / ml, DTT to a final concentration of 5mM, PMSF to a final concentration of 0.8m...

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Abstract

A new Der p1 nano-vaccine for the treatment of lung cancer and its preparation method and application, the method is: first clone and express the Der p1 protein in Escherichia coli by genetic engineering technology, and then use biodegradable PGLA nanoparticles as the package Materials Preparation PLGA-dust mite allergen Der p1 (PLGA-Der p1) nano-vaccine is the product of the present invention. A new Der p1 nano-vaccine for the treatment of lung cancer disclosed by the present invention has good anti-cancer function and significant therapeutic effect on lung cancer. .

Description

technical field [0001] The invention relates to the field of new vaccine inventions, in particular to a new Der p1 nano vaccine for treating lung cancer and its preparation method and application. Background technique [0002] Lung cancer has become the first in the incidence of all cancers in the world. Its five-year survival rate is about 15%. About 30-40% of lung cancer patients have bone metastasis. Once bone metastasis occurs, the median survival time is only 7 months. In 2012, there were more than 1.63 million new cases of lung cancer and 570,000 deaths in the United States. Lung cancer has also surpassed liver cancer to become the number one high-incidence cancer in my country, and its morbidity and mortality are increasing most rapidly, increasing at a rate of 13% per year. At present, lung cancer occupies the first place in the cancer incidence rate ranking (combined for men and women), among which it ranks first in the incidence rate of men, and always ranks in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/35A61K39/39A61P35/00
Inventor 左建宏刘志刚肖小军
Owner NANHUA UNIV
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