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A process for separating albumin from hemolyzed serum

An albumin and process technology, applied in the field of albumin separation, can solve the problems of large waste water treatment volume, high cost and high treatment cost, and achieve the effects of long freeze-drying time, high production efficiency and complicated operation.

Active Publication Date: 2022-04-15
GUANGZHOU RUITE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The salting-out method has the following disadvantages: the amount of ammonium sulfate is large, and 20-40 times the volume of pure water is required for dialysis desalination; time-consuming, 48 hours for dialysis, 24 hours for concentration, and 5 days for the entire process; low yield, generally 0.8-1 %; The amount of wastewater treatment is large, and dialysis desalination produces a large amount of wastewater, which contains high ammonium sulfate, and the treatment cost is high
The organic solvent precipitation method is the most commonly used E.J.Cohn method, but this method needs to be carried out at a low temperature of 0-5°C, which is time-consuming and laborious. The whole process takes 7 days, and the yield is not high, about 1%, and it is difficult to deal with environmental protection. Solvent recovery process, high cost
Moreover, the current albumin extraction method has high requirements for serum, which requires that the serum cannot be hemolyzed, which brings great restrictions on the preparation of albumin.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A process for separating albumin from hemolyzed serum:

[0031] 1) Accurately measure 1L of canine hemolyzed serum with a graduated cylinder;

[0032] 2) Accurately weigh 30 g of activated carbon with an electronic balance, add it to the serum and place it in -20°C for overnight freezing.

[0033] 3) Take out the frozen serum, place it at room temperature, and let it stand until the serum is completely frozen and thawed. At this time, the upper layer is an off-white solution mainly composed of albumin, and the lower layer is mainly composed of hemoglobin, fibronectin and fatty acids. The dark red solution was extracted, and the upper layer solution was transferred to a new container. In the lower layer solution, add 2 times the volume of pure water, repeat the operation of adding activated carbon to freeze the upper layer solution twice, and combine the upper layer solutions obtained twice. Then, suction filtration was performed with 0.4 μm filter paper, and the filtr...

Embodiment 2

[0047] 1) Accurately measure 1L of canine hemolyzed serum with a graduated cylinder;

[0048] 2) Accurately weigh 50 g of activated carbon with an electronic balance, add it to the serum and place it in -20°C for overnight freezing.

[0049] 3) Take out the frozen serum, place it at room temperature, and let it stand until the serum is completely frozen and thawed. At this time, the upper layer is an off-white solution mainly composed of albumin, and the lower layer is mainly composed of hemoglobin, fibronectin and fatty acids. The dark red solution was extracted, and the upper layer solution was transferred to a new container. In the lower layer solution, add 2 times the volume of pure water, repeat the operation of adding activated carbon to freeze the upper layer solution twice, and combine the upper layer solutions obtained twice. Then, suction filtration was performed with 0.45 μm filter paper, and the filtrate was collected to obtain solution 1.

[0050] 4) The solutio...

Embodiment 3

[0063] 1) Accurately measure 1L of canine hemolyzed serum with a graduated cylinder;

[0064] 2) Accurately weigh 40 g of activated carbon with an electronic balance, add it to the serum and place it in -20°C for overnight freezing.

[0065] 3) Take out the frozen serum, place it at room temperature, and let it stand until the serum is completely frozen and thawed. At this time, the upper layer is an off-white solution mainly composed of albumin, and the lower layer is mainly composed of hemoglobin, fibronectin and fatty acids. The dark red solution was extracted, and the upper layer solution was transferred to a new container. In the lower layer solution, add 2 times the volume of pure water, repeat the operation of adding activated carbon to freeze the upper layer solution twice, and combine the upper layer solutions obtained twice. Then, suction filtration was performed with 0.5 μm filter paper, and the filtrate was collected to obtain solution 1.

[0066] 4) The solution...

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Abstract

The invention discloses a process for separating albumin from hemolyzed serum, which is characterized in that it comprises the following steps: (1) adding activated carbon to the hemolyzed serum, freezing, and then standing at normal temperature until the serum is completely frozen-thawed and stratified, Extract the upper layer solution, filter and collect filtrate A; (2) Gel chromatography the filtrate A, collect the eluent to obtain solution B; (3) add carboxylated ferric oxide nano-magnetic beads to solution B, and after incubation at room temperature, Filtration and separation to obtain solution C; (4) ultrafiltration and concentration of solution C to obtain protein concentrate D; (5) virus inactivation and drying of protein concentrate D to obtain albumin powder. The raw material cost of the method is low, and the prepared albumin has good purity and high safety.

Description

technical field [0001] The invention relates to a process for separating albumin from hemolyzed serum. Background technique [0002] Serum protein is also called albumin. A class of small molecular weight, spherical, water-soluble proteins. Mainly found in mammals, bacteria, fungi and plants. Ovalbumin, lactalbumin, soy albumin, and serum albumin all fall into this category. Ovalbumin is an important protein in egg white, accounting for about 70% of the total protein of egg white. Serum albumin is the most abundant protein in serum, accounting for more than 50% of total serum protein, and it does not contain sugar. Serum albumin has an important physiological function in the blood, which is related to the maintenance of normal blood osmotic pressure. It is the main regulator of the total blood osmotic pressure. The lack of this albumin may cause body edema. Another role of serum albumin is to participate in the transport of fatty acids as a carrier of fatty acids. Seru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K1/36C07K1/34C07K1/16
Inventor 李长路
Owner GUANGZHOU RUITE BIOTECH CO LTD
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