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Preparation method of recombinant tumor specificity antiapoptotic factors with activity and application of products thereof

A tumor-specific, apoptotic factor technology, which is applied in the field of biopharmaceutical preparation, can solve problems such as incapable of industrialization, high toxicity and side effects of drugs, and impossibility of large-scale application.

Active Publication Date: 2010-12-22
沈阳金石生物制药有限公司
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Problems solved by technology

[0007] (3) Although there are a large number of existing antineoplastic drugs, there are some with better curative effect, such as paclitaxel, arsenic preparations, etc., but most of the drugs have severe side effects, and drug resistance often occurs after repeated medication, which leads to tumor recurrence. It is very necessary to develop anti-tumor drugs with less toxic side effects and significant curative effect
Due to its high technical and equipment requirements, this method is only suitable for laboratory research and cannot be applied on a large scale, so it cannot be industrialized

Method used

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  • Preparation method of recombinant tumor specificity antiapoptotic factors with activity and application of products thereof
  • Preparation method of recombinant tumor specificity antiapoptotic factors with activity and application of products thereof
  • Preparation method of recombinant tumor specificity antiapoptotic factors with activity and application of products thereof

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Embodiment Construction

[0037] (2) Fermentation, high-efficiency expression, dissolution, renaturation and purification of GST-Apoptin fusion protein, specifically:

[0038] (2.1) Activation, fermentation and recovery of inclusion bodies of engineered bacteria Inoculate engineered strains stored at -70°C on nutrient agar plate medium containing AP, and culture at 37°C for more than 12 hours; pick a single colony and inoculate into AP-containing medium In LB liquid medium, cultivate in a shaker at 37°C for more than 12 hours, inoculate the bacterial liquid into the fermenter at a ratio of 1:10, and cultivate at 37°C; when the concentration of the bacterial liquid reaches 2.0OD, add IPTG inducer; induce Cultivate for 4 hours, and at the same time, add a small amount of filler solution continuously, and the ammonia water automatically adjusts the pH value to 7.0. The cells were collected by centrifugation, suspended in phosphate buffer (PB, pH 7.2), and subjected to high-pressure homogenization to crush...

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Abstract

The invention provides a preparation method of recombinant tumor specificity antiapoptotic factors with activity and an application of products thereof, and the key points are as follows: (1) gene segments of Apoptin are directly obtained in an artificial synthesis method, and a prokaryotic expression vector pGEX-A comprising GST-Apoptin gene is constructed; (2) GST-Apoptin fusion protein is fermented, efficiently expressed, dissolved, renatured and purified; and (3) the GST-Apoptin fusion protein is chemically modified, thus obtaining the recombinant tumor specificity antiapoptotic factors with the activity, which can induce apoptosis of tumor cells and are applied to anti-tumor therapy. The invention has the beneficial effects of having efficient expression, good stability, high purity and high activity in Escherichia coli and being suitable for goingtoscale and industrialization.

Description

technical field [0001] The invention relates to a preparation method and application of biopharmaceuticals, especially a preparation method of an active recombinant tumor-specific apoptosis factor and the application of the product obtained by the method in anti-tumor treatment. Background technique [0002] Cancer treatment is a worldwide problem. Cancer biological treatment is an important supplement to surgery, radiotherapy, chemotherapy and other methods. Finding more effective biological agents has become one of the key research fields in the 21st century. At present, the biological agents for tumor treatment that have been marketed at home and abroad mainly include interleukin-2 (IL-2), interferon (IFN) and monocyte colony-stimulating factor (GSM). There are few varieties, and they are all immune enhancers or modulators, so the research and development of new biological agents with therapeutic effects has important social and economic significance. [0003] Chicken an...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K1/107A61K38/17A61P35/00
CPCY02A50/30
Inventor 赵洪礼
Owner 沈阳金石生物制药有限公司
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