Production process for separating and purifying fiber linking proteins from pig blood

A technology for separation and purification of fibronectin, which is applied in the field of extraction of biological preparations, can solve the problems of environmental pollution, waste of protein resources, low utilization rate of pig blood resources, etc., and achieve the effect of low equipment requirements and simple process

Inactive Publication Date: 2015-09-30
ANHUI BAODI MEAT FOODS
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Problems solved by technology

At present, the number of pigs slaughtered in my country is increasing day by day, but the utilization rate of a large amount of pig blood resources after slaughter is low, and it is mainly used as feed additives or low-value products such as edible blood tofu, which not only causes waste of protein resources, but also causes serious damage. environmental pollution

Method used

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Embodiment Construction

[0016] The present invention will be further described below in conjunction with embodiment:

[0017] A production process for isolating and purifying fibronectin from pig blood, comprising the steps of:

[0018] 1. Separation of plasma and blood cells: take 500mL whole blood added with 6‰ sodium citrate anticoagulant, centrifuge at 3000r / min for 20min, and separate plasma and blood cells.

[0019] 2. Salting out: Add an equal volume of 0.0175mol / LNa to the upper plasma 2 HPO 4 -NaH 2 PO 4 Buffer solution (pH 6.3) and mix well, then add 280g of ammonium sulfate while stirring to make the saturation reach 20%, adjust the pH to 7.0 with 5% NaOH, put it in a refrigerator at 4°C for 2 hours to fully precipitate. After centrifugation at 3000r / min for 15min, the main component of the obtained precipitate is fibronectin.

[0020] 3. Dialysis and desalination: Dissolve the 2-step precipitate in PBS buffer (pH6.3), put it into a dialysis bag for dialysis, and change the dialysate ...

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Abstract

The invention relates to a production process for separating and purifying fiber linking proteins from pig blood. The production process comprises the following steps: (1) separating blood plasma and blood cells from whole blood; (2) salting out: charging an acidic Na2HPO4-NaH2PO4 buffer solution into an upper layer of plasma, uniformly mixing, charging ammonium sulfate while stirring the solution so that the saturation degree reaches 20 percent, adjusting pH to be alkaline, placing the mixed solution into a refrigerator, refrigerating the mixed solution for 1.5 to 2.5 hours, sufficiently precipitating, finally centrifuging the solution, and obtaining a main precipitation component, i.e., fiber linking proteins; (3) dialyzing and desalting: dissolving precipitates in the step (2) into an acidic PBS buffer solution, putting the mixed solution into a dialysis bag, carrying out the dialysis and concentration, thereby obtaining a crude product of the fiber linking proteins; and (4) purifying: carrying out the affinity chromatography for an upper sample of the fiber linking protein crude product obtained in the step (3) by virtue of a gelatin sepharose-4B column, and eluting by virtue of a specific volume to obtain the purified fiber linking proteins. The production process is simple, low in requirement on needed equipment and suitable for industrialized production.

Description

technical field [0001] The invention relates to an extraction process of biological preparations, in particular to a production process for separating and purifying fibronectin from pig blood by a method combining centrifugation, salting out, dialysis and affinity chromatography. Background technique [0002] Fibronectin is a high-molecular glycoprotein widely present on the surface of various animal cells, extracellular fluid, connective tissue and most basement membranes. It has a variety of biological activities, such as participating in cell adhesion, differentiation, migration, proliferation, etc. , plays an important role in maintaining vascular integrity and permeability, wound healing, tumorigenesis and metastasis, blood coagulation and reproductive development. According to its distribution in the body, it is divided into two types: plasma type and cell type. The plasma type mainly exists in plasma and body fluids, also known as soluble fibronectin, and its content...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/36C07K1/34C07K1/30
Inventor 董浩张秀民胡瑞天陆银胜罗志良王雅静高振宏程榆茗
Owner ANHUI BAODI MEAT FOODS
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