Preparation method of pig thymosin

A technology of pig thymus and thymus, which is applied in the field of preparation of pig thymosin, can solve the problems of poor use efficiency of thymus, thymosin polypeptide structure, activity damage, limiting the application effect and use range of thymosin, and achieves the effect of improving the application effect and expanding the use range.

Pending Publication Date: 2022-07-29
福建农业职业技术学院 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing porcine thymosin production technology lacks strict and systematic inspection of exogenous viruses on pig thymus, especially the lack of inspection of African swine fever virus, an emerging swine disease in my country, resulting in a relatively large virus safety risk in the prepared porcine thymosin
[0007] (3) Due to the limitations of the technical level, there may still be porcine-derived viruses that have not been discovered or understood.
The disadvantage of this existing thymosin manufacturing technology is that the

Method used

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  • Preparation method of pig thymosin
  • Preparation method of pig thymosin
  • Preparation method of pig thymosin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A preparation method of porcine thymosin, comprising the following steps:

[0040] 1) Select the thymus of healthy pigs, porcine thymus is free of bovine viral diarrhea virus, porcine parvovirus, swine fever virus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, foot-and-mouth disease virus, pseudorabies virus and African swine fever Virus contamination, remove the fascia, muscle, and adipose tissue on the surface of the pig thymus, wash it with water for injection, mince it by a meat grinder, mix the water for injection with the minced pig thymus, and homogenize it by a colloid mill to obtain a homogenate. ;

[0041] 2) The homogenate was broken by cells, incubated at 60° C. for 0.5 h in water, and then centrifuged to obtain centrifugation supernatant and centrifugation precipitate respectively. The centrifuged precipitate was further mixed with water for injection, homogenized by a colloid mill to obtain a homogenate, and the cells wer...

Embodiment 2

[0051] A preparation method of porcine thymosin, comprising the following steps:

[0052] 1) Select the thymus of healthy pigs, porcine thymus is free of bovine viral diarrhea virus, porcine parvovirus, swine fever virus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, foot-and-mouth disease virus, pseudorabies virus and African swine fever Virus contamination, remove the fascia, muscle and adipose tissue on the surface of the porcine thymus, wash it with water for injection, mince it in a meat grinder, mix the water for injection with the pH value of 1.5 adjusted by hydrochloric acid and the minced porcine thymus, The homogenate was obtained by homogenizing with a colloid mill.

[0053] 2) The homogenate is subjected to cell fragmentation (repeated freeze-thaw fragmentation method), incubated at 60° C. in water for 2 hours, and then centrifuged to obtain centrifugation supernatant and centrifugation precipitate respectively. The centrifuged pr...

Embodiment 3

[0062] A preparation method of porcine thymosin, which comprises the following steps:

[0063]1) Select the thymus of healthy pigs, porcine thymus is free of bovine viral diarrhea virus, porcine parvovirus, swine fever virus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, foot-and-mouth disease virus, pseudorabies virus and African swine fever Virus contamination, remove the fascia, muscle, and adipose tissue on the surface of the pig thymus, wash it with water for injection, mince it by a meat grinder, mix the water for injection with the minced pig thymus, and homogenize it by a colloid mill to obtain a homogenate. .

[0064] 2) The homogenate was broken by cells, incubated at 60° C. for 1 h, and then centrifuged to obtain centrifugation supernatant and centrifugation precipitate, respectively. The centrifuged precipitate was further mixed with water for injection adjusted to pH 2.0 by hydrochloric acid, homogenized by a colloid mill to obta...

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Abstract

The invention discloses a preparation method of pig thymosin, which comprises the following steps: 1) mixing water for injection or water for injection of which the pH value is adjusted to 1.5-6.5 by hydrochloric acid with minced pig thymus, and homogenizing by a colloid mill to obtain homogenate; 2) carrying out cell disruption and water insulation and heat preservation on the homogenate, and centrifuging and collecting a centrifugal supernatant; (3) carrying out tangential flow filtration on the harvested centrifugal supernate by adopting a filter membrane, and collecting permeate; 4) carrying out tangential flow filtration on the permeate step by step by using filter membranes with different interception pore diameters and molecular weights, and collecting the permeate to obtain a pig thymosin crude product in a required molecular weight range; 5) mixing the thymosin crude products in at least one molecular weight range in the step 4) to obtain a thymosin primary product; and 6) removing viruses from the pig thymosin primary product, adjusting the pH value and osmotic pressure, filtering and sterilizing to obtain the pig thymosin. According to the method, step-by-step tangential flow filtration is adopted, an accurate and effective pig thymosin separation and concentration technology and a small molecule purification technology are established, and the pig thymosin in the required molecular weight range can be prepared by selecting filter membranes with different cut-off pore diameters and molecular weights according to different technological processes.

Description

technical field [0001] The invention belongs to the field of biological products, in particular to a preparation method of porcine thymosin. Background technique [0002] The thymus is an important immune organ of animals and can secrete a variety of important hormones, such as: thymosin, thymus humoral factor, thymopoietin and thymicfactor. Among them, thymosin, also known as thymosin, is a group of biologically active polypeptides (compositions) secreted by thymic tissue epithelial cells. , The adjuvant therapy of various diseases with low cellular immune function plays an important role. [0003] In 1961, Miller and Archer found that the thymus gland plays an important role in the differentiation, maturation and immune activity of lymphocytes in mice and rabbits, respectively. The thymosin mixture was isolated from . In 1972, Golden isolated thymosin fraction 5 (TF5) from calf thymus, and obtained α by isoelectric focusing electrophoresis. 1-10 , β 1-5 , γ, a total o...

Claims

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Application Information

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IPC IPC(8): C07K14/575C07K1/34C07K1/14A61K38/22A61P37/04
CPCC07K14/57581A61K38/2292A61P37/04
Inventor 徐磊钟敏万培伟王均辉王舒宁杨慧刘毅发谢杼倢廖惠珍钟云钦王宗冬陈雷朱海侠黄瑜张渊魁
Owner 福建农业职业技术学院
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