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A kind of preparation method of red blood cell substitute

A red blood cell and substitute technology, which is applied in the field of blood preparation of red blood cell substitutes, can solve the problems of loss of antioxidant function, shortened validity period, and harsh conditions, and achieve the effects of enhanced antioxidant performance, extended shelf life, and lower temperature requirements

Active Publication Date: 2014-10-22
SHANXI LIFEGEN
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Problems solved by technology

The reason is that there is a series of balance relationships among many contents in red blood cells. After purification, this balance is completely destroyed, and the function is relatively simple; the most obvious example is the antioxidant system, superoxide dismutase (SOD) and peroxidation. Hydrogenase (CAT) is the most important enzyme in the antioxidant system. It has been determined that the activities of SOD and CAT before hemoglobin purification are 8,000-30,000U / g and 30,000-150,000U / g respectively; after purification, their activities are respectively Between 0 ~ 150U / g and 0 ~ 300U / g, the loss of its antioxidant function leads to the need for subsequent production to be carried out under low temperature conditions, the conditions are relatively harsh, the cost increases, and the validity period is correspondingly shortened

Method used

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Examples

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Effect test

example 1

[0017] Step 1. Centrifuge 300ml of anticoagulated blood at 3000rpm for 20min, discard the supernatant, resuspend the cells with 100ml of red blood cells and 400ml of normal saline; repeat the above steps 5 times;

[0018] Step 2. Dilute the red blood cells to 30 mg / ml (calculated as hemoglobin, the same below) with physiological saline, and wash the red blood cells with a 0.45 μm hollow fiber column until the plasma protein concentration is less than 0.5 μg / ml;

[0019] Step 3, the preparation concentration is the phosphate buffer saline (pH7.5) of 20mM; According to the volume ratio of 1 to 1, slowly add phosphate buffer saline to the erythrocyte solution under stirring condition, stir 5 hours to make the content in the erythrocyte released;

[0020] Step 4, wash with physiological saline with a hollow fiber column with a pore size of 0.1 μm, and the filtrate is the red blood cell content;

[0021] Step 5, adjust the concentration of hemoglobin to 30 mg / ml (pH7.8), according...

example 2

[0024] Step 1. Centrifuge 300ml of anticoagulated blood at 4000 rpm for 15min, discard the supernatant, take 100ml of red blood cells and resuspend the cells with 400ml of normal saline; repeat the above steps 10 times;

[0025] Step 2, preparation concentration is the phosphate buffer saline (pH8.0) of 20mM; According to the volume ratio of 0.85 to 1, slowly add phosphate buffer saline to the erythrocyte solution under stirring condition, stir 10 hours to make the content in the erythrocyte released;

[0026] Step 3, wash with physiological saline with a hollow fiber column with a pore size of 0.1 μm, and the filtrate is the red blood cell content;

[0027] Step 4, adjust the concentration of hemoglobin to 50mg / ml (pH8.5), add superoxide dismutase and catalase, make the activity of two kinds of enzymes in the solution be respectively 30,000 U / ml and 100,000 U / ml ;

[0028] Step 5, according to the molar ratio that hemoglobin and malondialdehyde are 1 to 40, stir and add the...

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Abstract

A preparation method of a red blood cell substitute, the present invention is characterized in that the step of purifying the red blood cell content is canceled, and the red blood cell content is used to directly polymerize hemoglobin; compared with the prior art: the red blood cell substitute prepared by the present invention has stronger anti- Oxidation performance, so the shelf life is extended by more than three times; at the same time, the relatively complete function of red blood cells is retained; due to the reduction of the steps of hemoglobin purification, the requirements for temperature conditions in the preparation are reduced, the preparation process is simple, the production cycle is shortened, and the cost is reduced. Production cost; observed through animal contrast experiments, the prepared product has the effect of reducing adverse reactions; the preparation method of the present invention is suitable for preparing red blood cell substitutes by means of polymerized hemoglobin.

Description

technical field [0001] The invention belongs to the technical field of biological products, and in particular relates to a red blood cell substitute prepared from human or animal blood. Background technique [0002] Blood transfusion is a very important medical method in clinical operations, disaster relief, terrorist attacks and field rescue. At present, people mainly rely on people to donate blood to meet the needs. Due to the complex blood type of human beings, matching must be carried out during blood transfusion, and the storage period of blood is short, transportation is inconvenient, the source is limited, and there are potential risks of virus infection such as AIDS and hepatitis B. Limited, especially heavy use in emergencies. In recent years, with the continuous increase of blood demand, it is difficult to meet the increasing demand only by human blood donation. Therefore, the development of safe and effective blood substitutes is of great significance to alleviat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/805A61K35/14
Inventor 陈超严坤平朱宏莉
Owner SHANXI LIFEGEN
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