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Preparation method of sheep peripheral blood lymphocyte transfer factor solution and injection

A technology of lymphocytes and transfer factors, which is applied in the preparation methods of peptides, cytokines/lymphokines/interferons, blood/immune system cells, etc., to achieve the effect of increasing production and promoting a wide range of applications.

Pending Publication Date: 2022-08-05
商丘美兰生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, it is also necessary to consider whether the collected spleen is infected with exogenous microorganisms, which has certain risks.

Method used

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  • Preparation method of sheep peripheral blood lymphocyte transfer factor solution and injection
  • Preparation method of sheep peripheral blood lymphocyte transfer factor solution and injection
  • Preparation method of sheep peripheral blood lymphocyte transfer factor solution and injection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A preparation method of a sheep peripheral blood lymphocyte transfer factor solution, comprising the following steps:

[0026] Step 1: Preparation of Lymphocyte Seed Suspension:

[0027] (1) Collection of fresh sheep blood: Donor sheep should meet the current "Veterinary Pharmacopoeia of the People's Republic of China" on animal standards for production and inspection. After the sheep is sterilized, insert the blood collection needle into the sheep jugular vein, and aseptically collect blood into the blood collection bottle. Anticoagulation is available.

[0028] (2) Cell sedimentation and separation: Add 1 volume of anticoagulant physiological saline to the above anticoagulant and mix well, then dispense into each separatory funnel (1L), and settle naturally on the ultra-clean workbench for 24 hours . After 24 hours, flip the switch of the separatory funnel to remove the red blood cells at the bottom of the separatory funnel.

[0029] (3) Preparation of lymphocyte ...

Embodiment 2

[0051] For a transfer factor injection, the transfer factor solution obtained in Example 1 (obtained after culturing with optimal culture conditions) is sterile filtered using a 0.22 μm filter element. Then add mannitol and normal saline, mannitol: normal saline: transfer factor solution according to the volume ratio of 2:2:6 to prepare a transfer factor solution with a polypeptide content of not less than 1.5mg / ml and a ribose content of not less than 45ug / ml for injection liquid.

Embodiment 3

[0053] 1. Quality standard detection of transfer factor injection

[0054] According to the current appendix of "Veterinary Pharmacopoeia of the People's Republic of China", it should be aseptically grown. According to the determination method of polypeptide content in the quality standard of transfer factor oral liquid in the 2017 edition of "Veterinary Drug Quality Standard Biological Products Volume", we prepared three batches of transfer factor injection, and three batches of reference substances (purchased from Qingdao Yibang Bioengineering Co., Ltd. The company's pig spleen transfer factor injection) has been tested for polypeptide content. The content of the transfer factor polypeptide prepared by the present invention is not less than 1.5 mg per milliliter, which meets the requirements of veterinary drug quality standards, and the polypeptide content is higher than that of the reference substance (see Table 4 for the results). ).

[0055] Table 4 Detection and compari...

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Abstract

The invention provides a preparation method of a sheep peripheral blood lymphocyte transfer factor solution, which comprises the following steps: collecting fresh sheep blood, carrying out anticoagulation treatment, and carrying out cell sedimentation and separation; collecting a lymphocyte layer, and diluting the collected lymphocytes with Hank's liquid to prepare a lymphocyte seed suspension for later use; performing cell suspension culture on the lymphocyte seed suspension to obtain lymphocyte liquid; the lymphocyte fluid is subjected to cell disruption, the disrupted cells are filtered and ultra-filtered through a microfiltration membrane and an ultra-filtration membrane respectively, and ultra-filtrate is the obtained transfer factor solution; the invention also provides an injection which is prepared by performing sterile filtration on a transfer factor solution and then adding mannitol and normal saline. According to the method, sheep leukocytes are used as a culture medium, a cell suspension culture process is adopted for enlarged culture, the prepared transfer factor solution, namely the injection, can be suitable for large-scale production, and the yield is greatly improved.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a preparation method and an injection of a sheep peripheral blood lymphocyte transfer factor solution. Background technique [0002] Transfer factor (TF) is a class of small molecule dialyzable substances released by immune-learning T lymphocytes in animal protein cells. It is soluble in water and can induce the activation of immune cells and enhance immunity. The increase and decrease of cellular immune function and humoral immune function have two-way immunomodulatory effects, and have the characteristics of non-toxicity, non-antigenicity, non-allergic, non-pyrogenic and non-species specific, and their activity is not affected by trypsin, ribonucleic acid. destroyed by enzymes. Transfer factor has the advantages of safe use, rapid action, no antigenicity, and can significantly improve the prevention and control effect of animal diseases as an immune stim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/52C07K1/34C12N5/078A61K38/19A61K9/08A61P37/04A61P37/02
CPCC07K14/52C12N5/0634A61K9/08A61K9/0019A61P37/04A61P37/02A61K38/00C12N2509/00Y02A40/70
Inventor 王新港姬星宇张先锋李自波韩云涛丁丽萍周欣张真真张蒙蒙王彦辉
Owner 商丘美兰生物工程有限公司
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