Method for purifying and preparing superoxide dismutase in rabbit blood

A technology of superoxide and dismutase, applied in the field of biological protein, can solve the problems of complex process, serious environmental pollution, high cost, etc., and achieve the effect of simple operation and high purity

Inactive Publication Date: 2015-11-11
青岛康大食品有限公司 +1
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Problems solved by technology

The disadvantage of this process is that the process is complicated, the cost is high, and organic solvents such as chloroform an

Method used

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Embodiment Construction

[0018] The technical solution of the present invention will be described in detail below through examples.

[0019] The method for purifying and preparing superoxide dismutase in rabbit blood mainly comprises the following process steps:

[0020] 1. Add 50L of fresh rabbit blood to 4.0% sodium citrate (sodium citrate) at a ratio of 16:1, and stir well.

[0021] 2. Continuous flow centrifuge, the centrifugal force is about 400g. The centrifuged supernatant is plasma, and the precipitate is cells such as blood cells. Collect the precipitate.

[0022] 3. Wash the collected precipitate with 4 times normal saline, centrifuge at 4000 rpm in continuous flow, and repeat three times to remove hemoglobin. Then an equal volume of deionized water was stirred for hemolysis.

[0023] 4. Add an equal volume of 95% pre-cooled ethanol and 0.2 volume of pre-cooled chloroform to the hemolysis. After stirring for 20 minutes, let it stand for 1 hour, then centrifuge at 4000 rpm in continuous ...

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Abstract

The invention belongs to the technical field of biological proteins, and particularly relates to a method for purifying and preparing superoxide dismutase in rabbit blood. The method mainly includes technological steps of adding anticoagulants into the rabbit blood and centrifugally collecting precipitates; washing the precipitates by the aid of normal saline, centrifuging the precipitates and removing hemoglobin; stirring and dissolving the blood by the aid of deionized water with equal volumes; adding 95% of precooled ethyl alcohol with the equal volumes and precooled chloroform with 0.2-times volume into the dissolved blood; stirring and centrifuging the blood and acquiring supernatants; heating the supernatants until the temperatures of the supernatants reach 60 DEG C, preserving heat for 15 minutes, quickly cooling the supernatants until the temperatures of the supernatants reach the room temperature, centrifuging the supernatants and acquiring second supernatants; cooling the second supernatants in normal saline, adding precooled acetone into the second supernatants, stirring the second supernatants, acquiring second precipitates and dissolving the second precipitates by the aid of deionized water; carrying out desalination and chromatography on the second precipitates; washing the second precipitates twice by the aid of HAC-NaAC liquor, then dissolving the second precipitates again by the aid of PBS liquor and filtering the second precipitates by the aid of 0.45 micrometer filter paper; carrying out ion exchange chromatography on the second precipitates; carrying out ultra-filtration on the second precipitates; centrifuging SOD eluent under the condition of temperature of 4 DEG C, acquiring third supernatants which are SOD stock liquid. The method has the advantages that the method is easy to implement, and the superoxide dismutase extracted by the aid of the method is high in purity.

Description

technical field [0001] The invention belongs to the technical field of biological proteins, and in particular relates to a method for purifying and preparing superoxide dismutase in rabbit blood. Background technique [0002] Rabbit blood contains high nutritional value and can be processed into a variety of products for food, medicine, or as animal feed for livestock and poultry. Rabbit blood can extract a variety of biological drugs and biochemical reagents, such as medical serum, serum antigen, thrombin, leucine, peptone, superoxide dismutase, etc. Rabbit blood has high medical value, but the current rabbit blood extraction technology is limited to experiments Laboratory application, unable to achieve purification and extraction, and cannot be industrialized extraction and production, there are certain limitations in the application of rabbit blood. [0003] Superoxide dismutase Orgotein (SuperoxideDismutase, SOD), alias liver protein, abbreviation: SOD. SOD is an activ...

Claims

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Application Information

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IPC IPC(8): C12N9/02
CPCC12N9/0089C12Y115/01001
Inventor 张姝倩葛安山孙莎莎陈宴霞宋玉龙
Owner 青岛康大食品有限公司
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