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High-efficiency producing method for purified porcine leukocyte interferon by employing porcine spleen

A leucocyte interferon and white blood cell technology, which is applied in the field of efficient production of refined porcine leukocyte interferon, can solve the problems of low biological activity, difficulty in collecting pig blood, and poor clinical treatment effect

Inactive Publication Date: 2014-07-16
北京紫辰医药生物技术研究所
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Because pig blood is difficult to collect, it is not easy to store in large quantities, and the product has not been purified and extracted, and contains a large number of harmful components such as cell tissue, culture medium and other unknown pathogenic microorganisms, resulting in low biological activity (only 10,000-20,000 interferon per ml active units), the clinical effect is poor

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  • High-efficiency producing method for purified porcine leukocyte interferon by employing porcine spleen
  • High-efficiency producing method for purified porcine leukocyte interferon by employing porcine spleen

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Embodiment Construction

[0007] The specific implementation method of using pig spleen to efficiently produce refined porcine leukocyte interferon is as follows:

[0008] 1. Isolation of white blood cells from porcine spleen

[0009] (1) Aseptically collect fresh spleens from healthy pigs and store them frozen below -10°C for later use;

[0010] (2) Melt in a sterile water bath at room temperature during use, peel off the fat and connective tissue on the surface of the spleen, and grind it into 1-2mm pieces with a tissue grinder 3 pieces;

[0011] (3) Add pH 6.8 phosphate buffer containing gentamicin 40 units / ml and kanamycin 150 μg / ml for washing, centrifuge at 2000 rpm at 4°C for 15 minutes, and collect the cell tissue precipitate;

[0012] (4) Add 0.25% trypsin, and digest at 37°C for 30 minutes;

[0013] (5) Wash twice with phosphate buffer saline without antibiotics, filter through a stainless steel filter, centrifuge at 2000 rpm at 4°C for 15 minutes, and collect white blood cell precipitates...

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Abstract

The invention discloses a high-efficiency producing method for purified porcine leukocyte interferon by employing a porcine spleen and belongs to the technical field of biological pharmacy. The method includes steps of collecting fresh porcine spleens and cryopreserving the porcine spleens below -10 DEG C, melting, crushing, washing and digesting the frozen porcine spleens to separate leukocytes during usage, adding a Newcastle disease virus, carrying out an inductive fermentation culturing process in a fermentation tank, carrying out a separation process and a virus inactivating process to obtain a crude porcine leukocyte interferon, carrying out an ion-exchange process and a high-pressure reverse direction chromatography purification process to obtain a purified porcine leukocyte interferon original fluid, carrying out a vacuum freeze drying carrying out a vacuum freeze drying preparation process to obtain a purified porcine leukocytes interferon powder injection. The purified porcine leukocytes interferon belongs to a pure natural biological product, has effects of antivirus activity and increasing body immunity, and can be used for preventing and curing a plurality of viral infectious diseases such as swine fever, porcine reproductive and respiratory syndrome and diarrhea, etc. The purified porcine leukocytes interferon is produced with a raw material being the porcine spleens instead of porcine blood. The method is simple in technology, good in product quality and low in cost, and develops a new approach for producing the porcine leukocyte interferon.

Description

technical field [0001] The invention relates to a method for efficiently producing refined porcine leukocyte interferon by using pig spleen as a raw material, and belongs to the technical field of biopharmaceuticals for deep processing pig spleen by adopting cell separation technology, tissue culture technology, induced fermentation technology and preparation technology. Background technique [0002] Porcine leukocyte interferon is a pure natural cytokine with broad-spectrum anti-virus, anti-tumor and immune-enhancing functions. It also has the characteristics of safe use, good therapeutic effect, and low production cost. It is clinically used for porcine viral hyperthermia Disease, plague, blue ear disease, diarrhea and other infectious diseases prevention and treatment, received satisfactory results. [0003] Most of the porcine leukocyte interferons currently sold and used in the market are crude products prepared by using porcine blood leukocytes as raw materials. Becau...

Claims

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Application Information

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IPC IPC(8): C12P21/00C07K1/20C07K1/18
Inventor 李广善傅李芳
Owner 北京紫辰医药生物技术研究所
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