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Trichinella spiralis and tumor cell associated antigen protein and preparation method and medical application thereof

A technology of antigenic protein and trichinella, which is applied in the preparation method of peptide, anti-tumor drug, medical component of antibody, etc., can solve the problem of unclear anti-tumor active ingredients, and achieve the effect of easy preparation and industrialization

Inactive Publication Date: 2010-03-17
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Trichinella spiralis and tumor cell associated antigen protein and preparation method and medical application thereof
  • Trichinella spiralis and tumor cell associated antigen protein and preparation method and medical application thereof
  • Trichinella spiralis and tumor cell associated antigen protein and preparation method and medical application thereof

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Effect test

Embodiment 1

[0033] 1. Determination of antigens associated with Trichinella spiralis and myeloma cells

[0034] Firstly, the soluble antigen of Trichinella spiralis and SP2 / 0 myeloma cell antigen were prepared, and the positive sera of the two antigens were prepared by immunizing BalB / C mice respectively. The soluble antigen of Trichinella spiralis and SP2 / 0 myeloma cell antigen were coated according to the routine operation steps and reacted with SP2 / 0 myeloma cell positive serum, Trichinella spiralis positive serum and healthy mouse serum respectively, and the OD490 value was measured by a microplate reader. Taking P / N≥3 as the judgment standard, ELISA results showed that Trichinella spiralis muscle larval antigen and SP2 / 0 myeloma cell antibody and SP2 / 0 myeloma cell antigen and Trichinella spiralis antibody had good immune cross-reactivity, and the obtained The best antibody titers were: myeloma cell hyperimmune serum 1:6400; trichinella hyperimmune serum 1:3200, and neither antigen r...

Embodiment 2

[0043] Determination of antigens associated with Trichinella spiralis and liver cancer cells

[0044] Firstly, the soluble antigen of Trichinella spiralis and Hepa1-6 liver cancer cell antigen were prepared, and the positive sera of the two antigens were prepared by immunizing BalB / C mice respectively. The soluble antigen of Trichinella spiralis and the antigen of Hepa1-6 liver cancer cells were coated according to the routine operation steps and reacted with the positive serum of Hepa1-6 liver cancer cells, the positive serum of Trichinella spiralis and the serum of healthy mice respectively. / N≥3 is the judgment standard, ELISA results show that there are related antigens between Trichinella spiralis and Hepa1-6 liver cancer cells, and the best antibody titers of cross-reaction obtained are: Hepa1-6 liver cancer cell antigen immune serum 1: 400; Trichinella worm antigen immune serum 1:800. The soluble crude antigen of Trichinella spiralis has protein bands in the molecular ...

Embodiment 3

[0046] Determination of antigens associated with Trichinella spiralis and breast cancer cells

[0047] Firstly, ELISA was used to confirm the existence of relevant antigens between Trichinella spiralis and MCF-7 breast cancer cells, and then SDS-PAGE and Western-blotting were performed to determine its molecular weight. There are related antigens that can produce immune cross-reaction; SDS-PAGE results show that there are at least 6 protein bands with similar molecular weight between the MCF-7 breast cancer cell antigen and the Trichinella spiralis antigen: 6 protein bands with similar molecular weight: They are 66.2~90 (2 pieces), 52, 45, 26~35 (2 pieces) kDa, the most obvious of which are the two protein bands of 52 and 45 kDa; Western-blotting results show that the soluble antigen of Trichinella spiralis The 97.4kDa protein molecular weight range can react with the hyperimmune serum of MCF-7 breast cancer cells. The relative molecular weights of the reaction bands are 97, 6...

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Abstract

The invention provides a trichinella spiralis and tumor cell associated antigen protein and a preparation method thereof. The molecular weight of the antigen protein is 33.211 ku, pI is 4.64, and theantigen protein comprises 284 amino acids; and the trichinella spiralis and the tumor cell associated antigen protein can generate strong specific and non-specific immune reaction to the tumor, and meanwhile, is easy for preparation and industrialization. The invention also provides medical application of the protein for preparing a medicine for treating the tumor.

Description

Technical field: [0001] The invention provides a trichinella spiralis and tumor-associated antigen protein 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-associated antigens can generate strong specific and nonspecific immune responses against tumors. Invention content: [0003] The invention provides a trichinella spiralis and tumor cell-associated antigenic protein, which is a novel protein substance and has medicinal activity. [0004] The invention also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/34A61K39/00A61P35/00
Inventor 张西臣张楠李建华宫鹏涛张景芝杨举
Owner JILIN UNIV