Sterilization method of protein A immuno-absorbent column

An immunoadsorption and sterilization method technology, which is applied in the field of protein A immunoadsorption column sterilization, can solve the problems of active ingredient protein A degradation and denaturation, loss of removal of pathogenic substances, etc., and achieve low production cost and high level of sterility assurance , Solve the effect that can not be sterilized

Active Publication Date: 2015-12-09
GUANGZHOU KONCEN BIOSCI
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] For perfusate products, the best sterilization methods are steam sterilization and radiation sterilization, and quite a few perfusate products on the market currently use these two methods. However, for protein A immunosorbent columns, due to such extreme conditions The treatment will lead to the degradation and denaturation of the active ingredient protein A in the adsorption column, thus losing the ability to remove pathogenic substances from the blood

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  • Sterilization method of protein A immuno-absorbent column
  • Sterilization method of protein A immuno-absorbent column
  • Sterilization method of protein A immuno-absorbent column

Examples

Experimental program
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Embodiment 1

[0031] The sterilization method of the present embodiment is as follows:

[0032] Take 5g of protein A adsorbent, add 5ml of buffer solution with ion concentration of 0.2M, pH3~10 (choose citric acid buffer solution for pH3~6, phosphate buffer solution for pH7~8, glycine-sodium hydroxide buffer solution for pH9~10 liquid), bottled and steam sterilized at 100°C for 60 minutes.

[0033] Then detect the adsorption performance of the adsorbent before and after sterilization as follows:

[0034] Wash the adsorbent with a large amount of water, take 4.4ml and pack it into the column, first equilibrate the column with 10 times the column volume of the equilibrium solution (1×PBS, pH7.4), and then use 10 times the column volume of the eluent (0.1M, Glycine at pH 2.5) to wash the column, and then equilibrate the column with 10 times column volume of equilibration solution. Take 17ml of plasma to pass through the column at a flow rate of 1.7ml / min. After passing through the plasma, fi...

Embodiment 2

[0037] The sterilization method of the present embodiment is as follows:

[0038] Take 5g of protein A adsorbent, add 5ml of citric acid buffer (or physiological saline) with an ionic strength of 0.2M and pH6.0, bottle it for steam sterilization, and the sterilization conditions are 121°C and 15min (equivalent to the standard sterilization time F0=15min).

[0039] Then detect the adsorption performance of the adsorbent before and after sterilization with the method of embodiment 1, the results are shown in figure 2 ,from figure 2 It can be seen that under the sterilization conditions of 121°C and 15min, compared with the non-sterilized group, the protein A adsorbent with normal saline as the medium lost a lot of activity after sterilization (the activity retention value was only 28% ), and the protein A adsorbent added with the buffer solution can restore the activity to a greater extent after sterilization (the activity retention value is restored to 70%).

Embodiment 3

[0041] The sterilization method of the present embodiment is as follows:

[0042]Take 5g of protein A adsorbent, add 5ml of citric acid buffer solution (or physiological saline) with an ionic strength of 0.2M and pH 6.0, bottle it for steam sterilization, and the sterilization conditions are 115°C and 30min (equivalent to the standard sterilization time F0=8min).

[0043] Then detect the adsorption performance of the adsorbent before and after sterilization with the method of embodiment 1, the results are shown in image 3 ,from image 3 It can be seen from the figure that under the sterilization conditions of 115°C and 30 minutes, compared with the non-sterilized group, the protein A adsorbent with normal saline as the medium had an activity retention value of 39% after sterilization, while adding buffer After sterilization of the protein A adsorbent, the activity retention value is 75%. Compared with Example 2, it can be seen that the reduced steam sterilization condition...

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Abstract

The invention relates to a sterilization method of a protein A immuno-absorbent column. The sterilization method comprises the following steps: adding a protein A immuno-absorbent to a buffer solution which is 4-7 in pH value, packing a column and sterilizing with steam. The method further comprises a step of adding a stabilizer to the buffer solution, wherein the stabilizer consists of the following components: an antioxidant, polyhydric alcohol, polysaccharide, a chelating agent, a thickening agent, polypeptide or protein. By virtue of the sterilization process, the activity of the absorbent can be protected after sterilization, and the final sterilization of the protein A immuno-absorbent column can be achieved; and the obtained protein A immuno-absorbent column product, compared with products obtained from a sterile process, is higher in sterility assurance level and is lower in production cost.

Description

technical field [0001] The invention relates to the fields of biological materials and blood purification, in particular to a method for sterilizing protein A immunoadsorption columns. Background technique [0002] Protein A immunosorbent columns are mainly used in the treatment of autoimmune diseases. Autoimmune disease refers to the disorder of the immune system of the human body, which regards its own substances as foreign antigens and produces a series of immune reactions, resulting in the production of a large number of autoantibodies and immune complexes in the body, which in turn destroys human tissues and organs. General term for disease. Including myasthenia gravis, systemic lupus erythematosus, rheumatoid arthritis, Guillain-Barré syndrome and more than 80 kinds of diseases. Due to the unclear etiology, there is still no effective drug treatment for this kind of disease. Using protein A immunoadsorption column to directly adsorb and remove autoantibodies and imm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L2/07B01D15/20B01D15/38
Inventor 陈校园张磊余波光张海珍李乐军
Owner GUANGZHOU KONCEN BIOSCI
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