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Preparation method of trichina and tumor related protein gene and medical application

A technology related to proteins and Trichinella spiralis, applied in botany equipment and methods, biochemical equipment and methods, gene therapy, etc., can solve problems such as unclear anti-tumor active ingredients

Inactive Publication Date: 2012-05-02
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Trichinella spiralis is a nematode that parasitizes humans and various animals. In recent years, studies have found that it has strong anti-tumor activity, but its anti-tumor active ingredients are still unclear

Method used

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  • Preparation method of trichina and tumor related protein gene and medical application
  • Preparation method of trichina and tumor related protein gene and medical application

Examples

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Effect test

Embodiment 1

[0028] 1. Western blotting method to detect the cross-reaction between anti-SP2 / 0 myeloma cell antigen serum and Trichinella antigen: the method is: use PBS as negative control, perform SDS-PAGE on Trichinella muscle larvae antigen, and then perform Western blotting to detect Antiserum, negative serum, PBS and antigen reactivity of Trichinella spiralis muscle larvae. Antiserum and negative serum need to remove cross-reactive antibodies with E. coli before use. The method for removing cross-reactive antibodies to E. coli is to remove cross-reactive antibodies from antiserum by co-incubating with E. coli lysate. Cultivate 100 mL of Escherichia coli XL1-Blue strain in LB medium to reach saturation. Bacteria were harvested by centrifugation at 5000 g for 10 min at 4°C. Bacteria were resuspended in 3ml bacterial suspension buffer, frozen and thawed several times, and the bacteria were disrupted by ultrasonic. Centrifuge at 12 000g / min at 4°C for 30 minutes, and transfer the super...

experiment example 1

[0036] 2. Western blotting detection of antigens associated with Trichinella spiralis and SP2 / 0 myeloma cells

[0037] Using PBS as a negative control, SDS-PAGE was performed on the antigenic protein of Trichinella spiralis, and then Western blotting was performed to detect the reactivity of antiserum, negative serum, PBS and the antigenic antigen of Trichinella spiralis muscle larvae. After Western blotting, it can be found that anti-SP2 / 0 myeloma cell antigen serum can specifically combine with Trichinella spiralis antigen and myeloma cell antigen, while negative serum cannot specifically combine with Trichinella spiralis antigen and myeloma cell antigen.

[0038] 3. Immunological screening of antigens associated with Trichinella spiralis and SP2 / 0 myeloma cells

[0039] The anti-SP2 / 0 myeloma cell hyperimmune serum was used for cross-immune screening of the Trichinella spiralis cDNA expression library to obtain the antigen associated with the Trichinella spiralis and SP2 / 0 ...

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PUM

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Abstract

The invention provides a trichina and tumor cell related protein gene. The preparation method of protein gene comprises the following steps of: determining the related antigen existing between a tumor cell antigen and trichina positive serum and between a trichina antigen and tumor cell positive serum by utilizing an enzyme-linked immunosorbent assay; and screening a trichina cDNA (complementary deoxyribonucleic acid) expression library by utilizing SP2 / 0 hyperimmuneserum to obtain the related protein gene. The trichina and tumor cell related protein gene is a novel protein gene substance; tumor cells are sensitive to a variant identification substance, and the trichina and tumor cell related protein gene can produce strong specific and nonspecific immunoreaction to tumors, and the trichina and tumor cell related antigen is easy for preparation and industrialization.

Description

technical field [0001] The invention provides a trichinella spiralis and tumor-associated protein gene and a preparation method thereof, and also provides a medical application of the protein, which belongs to the field of biopharmaceuticals. Background technique [0002] Trichinella spiralis is a nematode that parasitizes humans and various animals. Recent studies have found that it has strong anti-tumor activity, but its anti-tumor active ingredients are still unclear. It is reported that malignant tumor cells are more sensitive to allogeneic markers, and immune cells produced by alloantigens are resistant to malignant tumors. Trichinella spiralis and tumor cell-related gene proteins can produce strong specific and non-specific immune responses against tumors. Contents of the invention [0003] The invention provides a protein gene related to trichinella spiralis and tumor cells, which is a new protein gene material and has biological activity. [0004] The invention ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/10A61K48/00A61K39/00A61P35/00
Inventor 张西臣李建华任保彦宫鹏涛程柏淇张国才张楠杨举李赫陈玉江
Owner JILIN UNIV
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