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Fusion protein and composition for treating animal tumors

A fusion protein and composition technology, applied in the field of anti-tumor drugs, can solve problems such as high cost, complicated preparation process, and difficult quality control, and achieve the effects of inhibiting metastasis, simple preparation process, and improving quality of life

Active Publication Date: 2021-01-12
BEIJING KENUOKEFU BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This brings problems such as complex preparation process, difficult quality control, and high cost.

Method used

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  • Fusion protein and composition for treating animal tumors
  • Fusion protein and composition for treating animal tumors
  • Fusion protein and composition for treating animal tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1IL12

[0037] Example 1 Construction of IL12 expressing cells

[0038] The coding region of the canine IL12 gene was synthesized, including two subunits, IL12a (Genbank ID: NM_001003293) and IL12b (Genbank ID: NM_001003292), connected with T2A sequence in the middle, and the two ends of the synthesized gene had BamHI and XhoI restriction sites respectively Spot, then digest with BamHI and XhoI, the system is as follows: 5 μg of IL12 plasmid, 4 μL of digestion buffer, 1 μL of BamHI, 1 μL of XhoI, add water to a total volume of 40 μL, and let stand at 37°C for 12 hours. Take out the EP tube, add 4.4 μL of 10× sample buffer, run electrophoresis on 1% agarose gel, recover the IL12 gene fragment after electrophoresis, and set aside.

[0039] Digest the expression vector pLentis-CMV-MCS-IRES-PURO, and the system is as follows: plasmid 2 μg, enzyme digestion buffer 3 μL, BamHI 1 μL, XhoI 1 μL, add water to a total volume of 30 μL, and stand at 37°C for 12 hours. Take out the EP tube, add...

Embodiment 2

[0043] Example 2 GMCSF expressing cell construction

[0044] Synthesize the coding region of the canine GMCSF (Genbank number: NM_001003245) gene, with BamHI and XhoI restriction sites at both ends of the synthesized gene, and then digest with BamHI and XhoI. The system is as follows: 5 μg of GMCSF plasmid, digestion buffer 4 μL, BamHI 1 μL, XhoI 1 μL, add water to a total volume of 40 μL, and let stand at 37°C for 12 hours. Take out the EP tube, add 4.4 μL of 10× sample buffer, run electrophoresis on 1% agarose gel, recover the GMCSF gene fragment after electrophoresis, and set aside.

[0045] Digest the expression vector pLentis-CMV-MCS-IRES-PURO, and the system is as follows: plasmid 2 μg, enzyme digestion buffer 3 μL, BamHI 1 μL, XhoI 1 μL, add water to a total volume of 30 μL, and stand at 37°C for 12 hours. Take out the EP tube, add 3.3 μL of 10× sample buffer, run electrophoresis on 1% agarose gel, recover the carrier fragment after electrophoresis, and set aside.

...

Embodiment 3I

[0049] Example 3 Construction of IL2 expressing cells

[0050] Synthesize the coding region of the canine IL2 (Genbank number: NM_001003305) gene, with BamHI and XhoI restriction sites at both ends of the synthesized gene, and then digest with BamHI and XhoI. The system is as follows: IL2 plasmid 5 μg, digestion buffer 4 μL, BamHI 1 μL, XhoI 1 μL, add water to a total volume of 40 μL, and let stand at 37°C for 12 hours. Take out the EP tube, add 4.4 μL of 10× sample buffer, and perform electrophoresis on 1% agarose gel. After electrophoresis, the IL2 gene fragment is recovered for use.

[0051] Digest the expression vector pLentis-CMV-MCS-IRES-PURO, and the system is as follows: plasmid 2 μg, enzyme digestion buffer 3 μL, BamHI 1 μL, XhoI 1 μL, add water to a total volume of 30 μL, and stand at 37°C for 12 hours. Take out the EP tube, add 3.3 μL of 10× sample buffer, run electrophoresis on 1% agarose gel, recover the carrier fragment after electrophoresis, and set aside.

...

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Abstract

The invention relates to a fusion protein and composition for treating animal tumors. The fusion protein comprises three different gene segments. The fusion protein obtained according to the inventionhas a good inhibition effect on various animal solid tumors, and can shrink the tumors and even regress the tumors; metastasis of malignant tumors can be better inhibited; and the life quality of a patient is improved to a great extent.

Description

technical field [0001] The invention relates to the technical field of antitumor drugs, in particular to a fusion protein and composition for treating animal tumors. Background technique [0002] Tumor is a new organism formed by the body under the action of various carcinogenic factors when a certain cell in a local tissue loses its normal regulation of its growth at the gene level, resulting in abnormal clonal proliferation. Because these new organisms are mostly space-occupying block-like protrusions, they are also called neoplasms. [0003] In recent years, many new methods have appeared in tumor treatment. At present, the effective treatment methods are radiotherapy, chemotherapy, surgery, immunotherapy and so on. However, there are still problems such as serious adverse reactions of radiotherapy and chemotherapy, high risk of surgical treatment, and poor effect of immunotherapy on solid tumors. [0004] In the pending Chinese patent application No. CN201810104146.9 s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K38/20A61K38/19A61P35/00
CPCC07K14/55C07K14/5434C07K14/535A61P35/00A61K38/00C07K2319/00C12N2740/16043A61K38/2013A61K38/208A61K38/193A61K2300/00
Inventor 张晋宇
Owner BEIJING KENUOKEFU BIOTECH CO LTD
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