Production technology of high purity yak serum albumin

A serum albumin and production process technology, applied in the preparation method of peptides, organic chemistry, peptides and other directions, can solve the problems of time-consuming, laborious, flammable ethanol, and high content of impurity proteins, and achieve easy process operation, high reliability and high reliability. The effect of safety and simple preparation process

Inactive Publication Date: 2016-03-09
QINGDAO ARCTIC YAK BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The salting-out method is the most traditional production method. The principle of this method is relatively simple, but ammonium sulfate is corrosive to the equipment. The whole process is time-consuming, and the amount of ammonium sulfate is large. The production rate is generally 0.8~1.0%, which leads to low production efficiency and high cost of this method; the use of cold ethanol precipitation method also has some shortcomings: ethanol is a non-specific precipitant, although the production efficiency of BSA is remarkable, the product purity is not very high , as an organic solvent, ethanol may also cause protein aggregation or denaturation. In order to avoid protein denaturation, it must be carried out in a low-temperature environment of -20°C. The investment in equipment and workshops is large, and ethanol is flammable, which brings safety hazards to production. One-way takes 7 days, energy consumption is huge, time-consuming and labor-intensive, and production is not easy to realize automation. The purity of BSA obtained by organic solvent precipitation is 90% to 95%, and the yield is about 10%. High price in the market reduces competitiveness
[0005] Yaks on the Qinghai-Tibet Plateau live in plateau areas above 3,000 meters above sea level. In order to adapt to the hypoxic environment in high-altitude areas, the number of red blood cells in their blood is much higher than that of ordinary bovine red blood cells. Simple application of traditional bovine serum albumin production methods will make the product The content of miscellaneous protein is high, and the traditional bovine serum albumin production method is not suitable for yak serum albumin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Take 56,000 mL of fresh yak blood from yaks that have undergone strict quarantine and are free from infectious diseases, and add it to 6,220 mL of 4.2% sodium citrate and 0.9% sodium chloride solution. After the blood is placed in a 3°C freezer for 10 hours, the plasma and red blood cells For stratification, use the siphon method to absorb the plasma, and the blood cells in the lower layer are used separately; place the above-mentioned separated plasma in a centrifugal system with a refrigeration device, set the temperature of the centrifugal system to 5°C, and turn on the centrifuge to centrifuge at a speed of 5000 rpm minutes, centrifuged for 50 minutes, and collected plasma. The plasma was then filtered with a 0.22 μm microporous membrane to obtain about 28,000 mL of purified plasma;

[0032]Put the refined plasma into the interlayer reaction tank, and heat the interlayer tank to 70°C; take 93.1g of sodium octanoate and 252g of sodium chloride, add them to 28000mL of...

Embodiment 2

[0034] Take 40,000 mL of fresh yak blood from yaks that have undergone strict quarantine and are free from infectious diseases, and add it to 4,450 mL of 4.2% sodium citrate and 0.9% sodium chloride solution. After the blood is placed in a cold storage at 8°C for 8 hours, the plasma and red blood cells Stratification, use the siphon method to absorb the plasma, and the blood cells in the lower layer are used separately; place the above separated plasma in a centrifugal system with a refrigeration device, set the temperature of the centrifugal system to 8°C, and turn on the centrifuge to centrifuge at a speed of 5000 rpm minutes, centrifuged for 45 minutes, and collected plasma. The plasma is then filtered with a 0.22 μm microporous membrane to obtain about 20,000 mL of purified plasma;

[0035] Put the refined plasma into the interlayer reaction tank, and heat the interlayer tank to 70°C; take 66.5g of sodium caprylate and 180g of sodium chloride, add them to 20000mL of purifi...

Embodiment 3

[0037] Take 50,000 mL of fresh yak blood from yaks that have undergone strict quarantine and are free from infectious diseases, and add it to 5,560 mL of 3.8% sodium citrate and 0.9% sodium chloride solution. After the blood is placed in a cold storage at 5°C for 8 hours, the plasma and red blood cells Stratify, use the siphon method to absorb the plasma, and use the lower layer of blood cells separately; put the above separated plasma in a centrifugal system with a refrigeration device, set the temperature of the centrifugal system to 10°C, turn on the centrifuge and centrifuge at a speed of 3000 rpm minutes, centrifuged for 60 minutes, and collected plasma. The plasma was then filtered with a 0.22 μm microporous membrane to obtain about 25,000 mL of purified plasma;

[0038] Put the refined plasma into the interlayer reaction tank, and heat the interlayer tank to 70°C; take 83.2g of sodium octanoate and 225.0g of sodium chloride, and add them to 25,000mL of purified water to...

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PUM

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Abstract

The invention discloses a production technology of high purity yak serum albumin. The production technology comprises the following steps: adding sodium citrate and sodium chloride into yak blood, separating plasma and erythrocyte, subjecting plasma to centrifugation, filtering and extraction, putting the plasma extract in an interlayer reaction pot, heating the pot, adding sodium caprylate, sodium chloride, and ethanol / methanol, slowing stirring, adding hydrochloric acid for twice to adjust the pH value, carrying out centrifugation, adding a precipitating reagent into the supernate, grinding the supernate into paste, stirring, freezing the paste for a night, carrying out centrifugation, collecting the precipitate, adding water to dissolve precipitate, adjusting the pH value by a sodium hydroxide solution, allowing the solution to stand still for a while, carrying out centrifugation, saving the supernate, filtering the supernate, inactivating the virus, and freeze-drying the supernate to obtain the finished product. The preparation technology is simple, the used materials are few, the technological procedure is short, the operation is easy, the product yield is high, the cost is low, and the product quality is good. The massive and automatic production can be easily realized. Moreover, the product is more reliable and safer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a production process of high-purity yak serum albumin. Background technique [0002] Bovine Serum Albumin (BSA) is a globulin in bovine serum, which contains 583 amino acid residues, a molecular weight of 66.43kDa, an isoelectric point of 4.7, a nitrogen content of 16%, and a sugar content of 0.08%. , the fat content is only 0.2%; albumin can be combined with various cations, anions and other small molecular substances. BSA is a biochemical reagent widely used in many disciplines of biology and in various fields of medicine and pharmacy. For example, it is used as a blocking reagent in western blot, Albumin in the blood mainly plays the role of maintaining osmotic pressure, pH buffering, carrier and nutrition; in the serum-free culture of animal cells, adding albumin can play the role of physiological and mechanical protection and carrier. [0003] There are many preparation methods...

Claims

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Application Information

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IPC IPC(8): C07K1/34C07K1/30
Inventor 许卫红
Owner QINGDAO ARCTIC YAK BIOLOGICAL TECH CO LTD
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