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Antitumor protein and application thereof

A protein and anti-tumor drug technology, applied in the field of anti-tumor drugs, can solve the problems of poor effect of solid tumors, high risk of surgical treatment, adverse reactions of radiotherapy and chemotherapy, etc., achieve good disease control, improve quality of life, and mild adverse reactions Effect

Pending Publication Date: 2019-08-09
BEIJING KENUOKEFU BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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.

Method used

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  • Antitumor protein and application thereof
  • Antitumor protein and application thereof
  • Antitumor protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1IL12

[0027] Example 1 Construction of IL12 expressing cells

[0028] The coding region of the canine IL12 gene was synthesized, including IL12α (Genbank ID: NM_001003293) and IL12β (Genbank ID: NM_001003292) two subunits, 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, and perform electrophoresis on 1% agarose gel. After electrophoresis, the IL12 gene fragment is recovered for use.

[0029] 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,...

Embodiment 2

[0033] Example 2 GMCSF expressing cell construction

[0034]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, and perform electrophoresis on 1% agarose gel. After electrophoresis, the GMCSF gene fragment is recovered for use.

[0035] 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, and run electrophoresis on 1% agarose gel. After electrophoresis, the carrier fragments are recovered a...

Embodiment 3I

[0039] Example 3 Construction of IL2 expressing cells

[0040] 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 run electrophoresis on 1% agarose gel. After electrophoresis, the IL2 gene fragment is recovered for use.

[0041] 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, and run electrophoresis on 1% agarose gel. After electrophoresis, the carrier fragments are recovered and set asi...

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PUM

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Abstract

The invention discloses an antitumor drug composition. The antitumor drug composition is characterized by comprising protein IL12, protein GMCSF and protein IL2, which achieves the technical effects of having a good inhibiting effect on tumors of all the stages and slight adverse reactions.

Description

technical field [0001] The invention relates to the technical field of anti-tumor drugs, in particular to an anti-tumor protein composition and its application. Background technique [0002] the tumor (tumor) is the body in various Carcinogen Under the action, one of the local tissues cell exist Gene Loss of normal regulation of its growth at the level, resulting in its clone The neogrowth formed by sexual dysplasia, because this neogrowth is mostly space-occupying block-like protrusions, also known as neoplasm (neoplasm). [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. Contents of the invention [0004] The purpose of the present invention is t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/20A61K48/00C12N15/26C12N15/24C12N15/12C12N15/63A61P35/00A61P35/02A61P35/04A61K38/19
CPCA61K38/193A61K38/2013A61K38/208A61K48/00A61P35/00A61P35/02A61P35/04C07K14/535C07K14/54C07K14/55A61K2300/00A61K38/19A61K38/20C12N15/63
Inventor 张晋宇
Owner BEIJING KENUOKEFU BIOTECH CO LTD
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