Serratia marcescens vaccine and preparation method and application thereof

A Serratia marcescens and bacterin technology, applied in the application field of medicine, can solve the problems of toxic side effects, retention, and insufficient curative effect, and achieve the effects of strong tumor inhibition, strong spleen activation, and stable yield

Active Publication Date: 2016-08-03
蔡剑前 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2002, He Xiaowen used the modified Telomere Repeat Amplification (TRAP)-silver staining method to detect the telomerase activity of Serratia marcescens preparations in 20 cases of bladder transitional cell carcinoma tumor tissues before and after perfusion, and found that Serratia marcescens The preparation can reduce the activity of telomerase in tumor tissue
[0005] However, many of the above-mentioned researches are not outstanding enough in curative effect, or have certain toxic and side effects, or cannot be industrialized due to the limitation of the expanded culture process of the strain, and have not developed into a real clinical drug so far, and have been stuck in the basic research stage. Seriously restricting the clinical application of Serratia marcescens in anti-tumor

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  • Serratia marcescens vaccine and preparation method and application thereof
  • Serratia marcescens vaccine and preparation method and application thereof
  • Serratia marcescens vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: the screening of bacterial strain

[0043] 1. Screening of starting strains

[0044] The strains are derived from Serratia marcescens preserved in the China General Microorganism Culture Collection and Management Center. The preservation number is: CGMCC1.0589. Under sterile conditions, the original strains are diluted 10 times and then spread on LB slant medium. Incubate at 37°C for 24h. Select a plate with about 20 colonies, pick a single colony with a large, clear boundary, and a milky white color that grows well as the starting strain for ultraviolet mutagenesis, and take its bacterial body in a triangular flask with normal saline and glass beads to make a sterile Bacteria suspension 500mL, 30 ℃, 250rpm water bath 4h, prepared into 10 6 CFU / mL of the mutagenized bacterial suspension, for later use.

[0045] 2. Determination of strain mutagenesis and lethality

[0046] Preheat the UV lamp for 25 minutes, take 10 mL of the mutagenic bacteria suspensi...

Embodiment 2

[0075] Embodiment 2: Optimization of preparation process of Serratia marcescens bacterin vaccine

[0076] During the research, it was found that the pH of LB agar medium, the time of slant culture, the temperature of slant culture, the pH of LB liquid medium, the inoculum size, the ventilation rate, the fermentation temperature and time and the rotation speed had a great influence on the growth and reproduction of bacteria. The preparation process of Serratia bacterin vaccine has been optimized, as follows:

[0077] 1. Screening of LB agar medium pH for spreading slant strains

[0078] According to the selected LB agar medium formula, the dosage of other components remained unchanged, and Serratia marcescens was cultured on LB agar medium with different pH at 37°C for 21 hours.

[0079] The results show that the bacteria can grow and reproduce on the agar medium with pH 6.0-8.0; because the bacteria will produce various acidic metabolites during the growth and metabolism proc...

Embodiment 3

[0113] Embodiment 3: the preparation of Serratia marcescens bacterin injection

[0114] The preparation method of Serratia marcescens injection is as follows:

[0115] (1) Slant-transfer strains: Serratia marcescens (strain SM-1 screened in Example 1) was inoculated on LB agar medium (pH 7.2) in a Kerner flask, and cultured at 37° C. for 21 hours. Take it out and store it in a -80°C refrigerator for later use.

[0116] (2) Cultivation and fermentation: Take 3 single colonies from the original slant strain of Serratia marcescens, mix them evenly, inoculate them on the LB liquid medium in the Erlenmeyer flask, and cultivate and ferment. The conditions are: LB liquid medium pH 7.2, 10% inoculum size, 75% aeration, temperature 37° C., 21 h, rotation speed 180 rpm.

[0117] (3) Bacteria collection, sterilizing and washing: before collecting bacteria, carry out the pure species inspection such as the morphological characteristics of Serratia marcescens, centrifuge at 4000-6000rpm ...

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Abstract

The invention discloses a serratia marcescens vaccine.The active ingredient of the serratia marcescens vaccine is a biological pure culture of serratia marcescens.The serratia marcescens is preserved in China General Microbiological Culture Collection Center on January 11, 2016 with the preservation number of CGMCC NO.11987.The invention further discloses a preparation method of the serratia marcescens vaccine.The serratia marcescens vaccine is prepared from the biological pure culture of the serratia marcescens through a proper separation method.According to the proper separation method, centrifugation is carried out at 4,000-6,000 rpm for 30-60 min, supernatant is discarded, sediment is inactivated, centrifugation is carried out again, supernatant is discarded, sediment is washed and centrifuged, and the serratia marcescens vaccine is obtained through preparation.The product produced through the preparation method is stable in yield, good in reproducibility and high in anti-tumor effect and spleen activity, and all other indexes meet the requirements of the quality standard.The serratia marcescens vaccine has obvious prevention and treatment effects on various solid tumors, and is an inactivated vaccine which is safe and free of toxic and side effects.

Description

technical field [0001] The invention relates to a Serratia marcescens vaccine and its preparation method and application, in particular to the preparation of the Serratia marcescens vaccine and its application in the preparation of drugs for preventing and / or treating tumors. Background technique [0002] According to WHO's "Global Cancer Report 2014", the number of cancer patients and deaths in the world increased disturbingly in 2012. In 2012, there were 14 million new cancer cases and 8.2 million deaths worldwide. It is predicted that global cancer cases will show a rapid growth trend, reaching 19 million in 2025 and 24 million in 2035. Nearly half of the new cancer cases occurred in Asia, most of which were in China, which ranked first in new cancer cases. [0003] According to the 2014 data from the China Cancer Registry Center, the number of cancer cases in my country accounts for 1 / 5 of the world, and the number of cancer deaths in China accounts for 1 / 4; 6 people ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/74A61P35/00C12R1/43
CPCA61K35/74C12N1/20C12N1/205C12R2001/43
Inventor 蔡剑前张忆军孟红陈雪梅孙永庆
Owner 蔡剑前
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