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Purification method of tetanus immune globulin and affinity chromatography filler of tetanus immune globulin

A technology of immunoglobulin and human immunoglobulin, applied in the field of human immunoglobulin purification, can solve the problem of lack of tetanus antibody enrichment, etc., achieve low possibility of clinical side effects, suitable for clinical use, and comprehensive utilization of plasma Enhanced effect

Pending Publication Date: 2022-07-12
HUALAN BIOLOGICAL ENG CHONGQING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention intends to provide a method for purifying tetanus immunoglobulin to solve the technical problem that the prior art lacks an effective method for enriching tetanus antibodies in plasma

Method used

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  • Purification method of tetanus immune globulin and affinity chromatography filler of tetanus immune globulin
  • Purification method of tetanus immune globulin and affinity chromatography filler of tetanus immune globulin
  • Purification method of tetanus immune globulin and affinity chromatography filler of tetanus immune globulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of chromatography packing

[0034] (1) Ligand preparation: The raw materials of the ligand include finished tetanus toxoid products, semi-finished products and stock solutions, which can be obtained through commercial channels. Among them, tetanus toxoid is the main functional component of tetanus vaccine. Use a 0.2 μm filter element to filter and concentrate the above raw materials, so that the tetanus toxoid titer reaches more than 5000 IU / ml. Dilute the titer to 1000-2000IU / ml with coupling buffer (0.2mol / L sodium bicarbonate+0.5mol / L sodium chloride, pH7.50-8.50) to obtain the ligand solution.

[0035] (2) Medium preparation: The cross-linked agarose-based medium is washed 3-5 times with a 1 mmol / L hydrochloric acid solution at 0-2° C. to obtain an activation medium.

[0036] (3) Ligand coupling: The washed activation medium is mixed with the ligand solution at a ratio of 1:1-1:100 (that is, 1ml of the activated medium is mixed with 1ml-100ml ...

Embodiment 2

[0038] Example 2: Purification of Tetanus Immunoglobulin

[0039] (1) Chromatography packing (layered medium) packing: Load the equilibrated microspheres into the chromatography column.

[0040] (2) Preparation of products: prepare an immunoglobulin solution containing tetanus antibodies, including raw plasma, cold gel-removed plasma, component II+III, component II, secondary precipitation and other precipitation lysates after filtration, component I, Component III, primary supernatant, etc. after ultrafiltration to remove ethanol, that is, components obtained in each step in the plasma production process can be used as raw materials. If it is in the precipitation state, it needs to be dissolved and filtered, and if the solution contains ethanol, it needs to be removed by ultrafiltration. The immunoglobulin solution containing tetanus antibody also includes the specific human immunoglobulin produced, the intravenous human immunoglobulin stock solution, and the semi-finished o...

Embodiment 10

[0054] Embodiment 10: get the precipitation of component II in the low-temperature ethanol method production process, use 6 times of the precipitation weight to dissolve in water for injection, use a deep filter stack and a terminal to filter the product in series with 0.2 μm after dissolving, and then follow the pH and conductivity of Example 6. The product solution before chromatography is required to be prepared according to the protein concentration, and then chromatography, ultrafiltration and preparation are carried out according to the method of Example 6 to obtain a finished product of tetanus human immunoglobulin.

[0055] The specific acquisition process of component II is as follows:

[0056] After the raw plasma is released from the warehouse, use 75vol.% ethanol solution to sterilize the surface of the plasma bag, then break the plasma bag, control the temperature to melt at 0°C, combine the plasma after melting, centrifuge the plasma using a centrifuge (centrifuga...

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Abstract

The invention relates to the technical field of purification of human immune globulin, in particular to a purification method of tetanus immune globulin and affinity chromatography filler of the tetanus immune globulin. The purification method of the tetanus immune globulin comprises the following steps: carrying out affinity chromatography treatment on a product by using a chromatographic column; the chromatographic column is filled with a chromatographic filler, and the chromatographic filler is coupled with an affinity substance for specifically binding with a tetanus antibody. The affinity chromatography filler comprises the following raw materials: a cross-linked agarose-based medium and ligands, wherein the ligands comprise a tetanus toxoid finished product, a tetanus toxoid semi-finished product and a tetanus toxoid stock solution. According to the technical scheme, the technical problem that a method for effectively enriching tetanus antibodies in plasma is lacked in the prior art can be solved. The tetanus human immune globulin produced by the process is lower in protein concentration and higher in purity, the comprehensive utilization rate of plasma is increased, and the market supply quantity of the tetanus human immune globulin is increased.

Description

technical field [0001] The invention relates to the technical field of human immunoglobulin purification, in particular to a purification method of tetanus immunoglobulin and an affinity chromatography filler. Background technique [0002] Tetanus is a disease caused by the infection of anaerobic bacteria Clostridium tetani. When the skin and mucous membranes are ruptured, Clostridium tetani can invade the human body through the ruptured site and multiply on the wound. Produce neurospastin and hemolytic toxin, such as improper disposal, can easily lead to death of patients. The current methods to prevent the occurrence of this disease are: first, thoroughly debride the wound of the exposed tetanus patient, and then inject a sufficient amount of tetanus antitoxin or tetanus human immune globulin or tetanus vaccine according to the injury. Since tetanus antitoxin can cause two types of allergic reactions, anaphylactic shock and serum sickness, and other adverse reactions in c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K1/22C07K1/14B01J20/286
CPCC07K16/1282B01J20/286Y02A50/30
Inventor 张宝献肖岚滕世超刘余江张建璀周康森
Owner HUALAN BIOLOGICAL ENG CHONGQING
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