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Method for preparing recombined apoptosis induction ligand relevant to human soluble tumor necrosis factor

A technology of tumor necrosis factor and apoptosis-inducing ligand is applied in the field of preparation of medicinal protein substances, which can solve the problems of drug resistance of tumor cells, inability to receive effective treatment for tumor patients, and strong toxic and side effects of normal cells, and achieve industrialization. production effect

Active Publication Date: 2005-11-16
深圳未名新鹏生物医药有限公司
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Problems solved by technology

However, the feedback data after decades of clinical application shows that although chemotherapy is currently the preferred anti-tumor regimen, most anti-tumor chemotherapeutic drugs have two serious problems: strong toxic side effects on normal cells and drug resistance of tumor cells sex, resulting in cancer patients not being able to receive effective treatment
At present, TRAIL is still in the stage of laboratory research, and there are no reports on pilot tests and industrialization.

Method used

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preparation example Construction

[0012] The preparation method of recombinant human soluble tumor necrosis factor-related apoptosis-inducing ligand of the present invention includes step a. Selecting engineering bacteria rsTRAIL96-281-pET15b-BL21(DE3)CGMCC No.0659 as a bacterial strain for fermentation and culture, also includes step: b .washing and ultrasonic crushing the bacterial cells obtained from the fermentation culture; c. performing salting-out treatment on the bacterial cells obtained from the crushing; d. performing chromatographic purification processing on the bacterial cells obtained from the salting-out treatment; e. Bacterial substances were subjected to buffer system replacement.

[0013] Said step b comprises sub-steps:

[0014] ①. Adding NaCl-containing Tris Cl [tris(hydroxymethyl)aminomethane hydrochloric acid] solution to the fermented thallus, and performing centrifugation to obtain thallus precipitation;

[0015] ②. Add EDTANa to the cell pellet 2 (disodium edetate) and DTT (dimercapt...

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Abstract

A process for preparing the wither inducing ligand (TRAIL) associated with recombinant human soluble tumor necrosis factor includes such steps as providing thue engizzeered bacterium rsTRAIL96-281-pET156-BL21 (DE3) (CGMCC No.0659), fermenting culture, washing, ultrasonic breaking, salting-out, chromatography and exchanging in buffering liquid system.

Description

technical field [0001] The invention relates to the preparation of medicinal protein substances, in particular to a method for preparing target proteins by using genetically engineered bacteria, in particular to a preparation method for recombinant human soluble tumor necrosis factor-related apoptosis-inducing ligands. Background technique [0002] Tumor is a common and frequently-occurring disease that seriously threatens human life, and its high fatality rate accounts for the second cause of all deaths. However, the feedback data after decades of clinical application shows that although chemotherapy is currently the preferred anti-tumor regimen, most anti-tumor chemotherapeutic drugs have two serious problems: strong toxic side effects on normal cells and drug resistance of tumor cells Therefore, cancer patients cannot receive effective treatment. While traditional chemotherapy drugs kill tumors, they "do not distinguish between the enemy and the enemy", have strong toxic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74C12P21/00
Inventor 李政海胡家华田宾李继东
Owner 深圳未名新鹏生物医药有限公司
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