Preparation method of pig thymosin

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...

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 heat treatment at 70-85°C has great damage to the polypeptide structure and activity of thymosin

Existing thymosin production technology has poor utilization efficiency for thymus. Only the centrifuged supernatant is used to prepare thymosin after centrifugation of the homogenate, and the centrifuged sediment is discarded.

In addition, the existing thymosin manufacturing technology lacks the osmotic pressure adjustment process, and it is impossible to obtain isotonic thymosin, which limits the application effect and scope of thymosin

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