Composite tumor gene vaccine taking bacterial nano magnetosome as carrier and preparation method thereof

A tumor gene and nano-magnetic technology, which is applied in the fields of biotechnology and tumor immunity, can solve the problems of incomplete research on the application conditions

Inactive Publication Date: 2011-04-27
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there are reports at home and abroad that it can be used as a method of gene delivery, the conditions for its application have not been thoroughly studied.

Method used

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  • Composite tumor gene vaccine taking bacterial nano magnetosome as carrier and preparation method thereof
  • Composite tumor gene vaccine taking bacterial nano magnetosome as carrier and preparation method thereof
  • Composite tumor gene vaccine taking bacterial nano magnetosome as carrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Transformation, extraction and purification of plasmids (see the second edition of "Modern Molecular Biology Experimental Technology" for specific methods, Peking Union Medical College Press)

[0041] Plasmids pSLC-E7-Fc, pEGFP-N1 and pGL4.17 (DNA) prepared by this method were used for subsequent experiments.

Embodiment 2

[0043] Preparation of compound tumor gene vaccine (BMP-PEI / pSLC-E7-Fc):

[0044] (1) Sterilize the pure bacterial nano-magnetosomes (gifted by Professor Li Ying of China Agricultural University) by irradiating with 15kGy gamma rays; measure the light absorption of bacterial nano-magnetosomes at 660nm according to the method of Nakamura, N. et al. Value (Nakamura, N., Mutsunaga, T.Highly sensitive detection of allergen using bacterialmagnetic particles: Biosensors.Anal Chem Acta, 1993, 281, 585-589) and according to 10D 660 = 172 μg bacterial nanomagnetosomes for quantification.

[0045] (2) The plasmid pSLC-E7-Fc (tumor gene vaccine) prepared in Example 1 was measured by ultraviolet spectrophotometry, and quantified (see the second edition of "Molecular Cloning" for specific methods, Science Press), and the tumor gene vaccine The triple anti-tumor vaccine pSLC-E7-Fc constructed for the inventor's laboratory, the size is 2.3kb (Liu R, Zhou C, Wang D, etal.Enhancement of DNA va...

Embodiment 3

[0049] Tumor Prevention Experiment of Plasmid pSLC-E7-Fc in Mice

[0050] Forty 6-week-old C57 female mice were randomly divided into 8 groups: pCMV group, pE7 group, pFc group, pSLC group, pE7-Fc group, pSLC-Fc group, pSLC-E7-Fc group and pSLC+pE7 group +pFc group, 5 animals per group. Immunization procedure: On the 10th day before TC-1 tumor cell inoculation, various plasmids were immunized with a gene gun. On day 0, 5 × 10 4 TC-1 cells. Observe the tumor emergence time. The result is as figure 1 As shown, within 60 days, 100% of the mice immunized with the pSLC-E7-Fc group did not grow tumors, while the mice immunized with all the control plasmids had full-length tumors; Mice also grow tumors in full.

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Abstract

The invention relates to a composite tumor gene vaccine taking a bacterial nano magnetosome as a carrier and a preparation method thereof. The range of a pH value of the optimal phosphate buffer system when the bacterial nano magnetosome is connected with the gene vaccine and magnetic field intensity and magnetic field acting time used by in-vitro transfection and in-vivo immunization are provided specifically, and treatment dosage range and immunization way are determined when the composite gene vaccine is used for treating a tumor model. A novel simple and convenient gene delivery mode is established, and a preparation condition and an immunization method of a new gene vaccine are provided for gene treatment.

Description

technical field [0001] The invention relates to the fields of biotechnology and tumor immunity, in particular to a compound tumor gene vaccine with bacterial nano magnetosomes as carriers and a preparation method thereof. Background technique [0002] Tumor gene therapy is a treatment that uses gene transfer technology to introduce foreign genes into host cells, directly repair or correct the structural and functional defects of tumor-related genes, or kill tumor cells by enhancing the host's immune defense function. Tumor DNA vaccine is the hotspot in tumor immune gene therapy research. At present, the methods of genetic immunization in the body mainly include direct intramuscular injection immunization, electric pulse, gene gun, etc. Each of these methods has advantages and disadvantages. Although direct intramuscular injection of immunization is simple, the amount of genes required is large, and the immune response elicited is effective. Electric pulses affect many fact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/48A61P35/00A61K47/59
Inventor 张叔人唐翌姝王冬梅周春霞马文波刘容枝
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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