A kind of genetic engineering recombinant trail fusion protein and its preparation method and application

A fusion protein and genetic engineering technology, applied in the field of genetic engineering recombinant TRAIL fusion protein and its preparation, can solve the problems of poor stability, short half-life of TRAIL, weak biological activity, etc., and achieve simple method, good anti-tumor or cancer cell activity, The effect of high purity
CN106084063BActive Publication Date: 2020-10-30JILIN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
JILIN UNIV
Publication Date
2020-10-30

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Abstract

The invention discloses a recombinant protein with improved stability and activity, obtained by using adenovirus fibrin fragments as a trimer motif and adopting TRAIL fusion expression and used as an antitumor therapy drug. The invention also provides a method for expression, purification and activity detection of the fusion protein. The method concretely comprises the following steps: 1, designing and constructing fusion gene, and carrying out low-temperature inducible expression by using an Escherichia coli system; 2, purifying the target protein by using a nickel column affinity chromatography technology, and removing an His label by using a thrombin digestion site to obtain the final target product; 3, detecting the killing activity of the fusion protein to tumor cells and the toxicity of the fusion protein to normal cells in vitro; 4, detecting the stability of the fusion protein under different conditions; and 5, detecting the in vivo antitumor activity of the fusion protein in a nude mouse model. Two adenovirus fibrin fragments are used as the TRAIL fusion protein of the trimer motif. The invention provides a method for simply and rapidly obtaining the high-stability TRAIL protein. The method can be applied to antitumor researches of the TRAIL protein. The abuses of poor stability and low activity of the TRAIL protein are solved.
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Description

technical field

[0001] The present invention relates to a fusion protein, its preparation method and application, in particular to a gene engineering recombinant TRAIL fusion protein, its preparation method and application. Background technique

[0002] Tumor necrosis factor-related apoptosis-inducing ligand (TNF related apoptosis-inducing ligand, TRAIL) is a member of the TNF superfamily, has a broad-spectrum anti-cancer function, and it is almost non-toxic to normal tissues and cells. The active TRAIL is in a trimer state, which is maintained by chelation between the zinc ion in the center of the trimer and the cysteine ​​on each monomer. However, TRAIL itself has short half-life, poor stability, and weak biological activity, and some studies have shown that the structural order of soluble TRAIL trimer will change after the introduction of tags, resulting in hepatotoxicity. Contents of the invention

[0003] The purpose of the present invention is to provide a genetical...

Claims

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