Method for low-temperature extraction of human serum albumin employing ethanol

A technology of human albumin and low-temperature ethanol, which is applied in the field of preparing human albumin, can solve the problems that the albumin production yield needs to be improved, the precipitation of extracted components is not refined enough, the component separation period is long, and the like, so as to shorten the separation time. Period, beneficial to comprehensive utilization, mild effect of mixed reaction

Inactive Publication Date: 2014-04-09
HUALAN BIOLOGICAL ENG CHONGQING
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Problems solved by technology

The problem of the low-temperature ethanol method is that the yield of albumin is low and the stability is poor, so this method needs to be improved to increase the yield of albumin
The c

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  • Method for low-temperature extraction of human serum albumin employing ethanol

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Embodiment 1

[0024] see figure 1 , the method for extracting human serum albumin with low-temperature ethanol, using human plasma as raw material, adopting low-temperature ethanol method and pressure filtration method to produce human serum albumin, the production operating pressure is less than 0.25MPa, comprising the following steps:

[0025] Step 1. Isolation of FI from human plasma:

[0026] Control the temperature of the fused human plasma at 0.0°C, adjust the protein concentration to 40g / L, adjust the solution conductivity to 12.5ms / cm, pH value to 6.80, ethanol concentration to 7.0% by volume, and control the temperature to -1.0°C After the precipitation is complete, use a filter press to filter under pressure, choose a deep layer filter plate with a pore size of no more than 0.9 μm (such as 0.9 μm), control the temperature of the filter press to below -1°C, and filter to separate FI was precipitated to obtain FI supernatant;

[0027] Step 2. Separation of FⅡ+Ⅲ from the supernatan...

Embodiment 2

[0037] see figure 1 , the method for extracting human serum albumin with low-temperature ethanol, using human plasma as raw material, adopting low-temperature ethanol method and pressure filtration method to produce human serum albumin, the production operating pressure is less than 0.25Mpa, including the following steps:

[0038] Step 1. Isolation of FI from human plasma:

[0039]Control the temperature of the fused human plasma between 2.0°C, adjust the protein concentration to 52.5g / L, adjust the solution conductivity to 13.1ms / cm, pH to 7.05, ethanol concentration to 8.5% by volume, and control the temperature to -2.0°C, after the precipitation is complete, use a filter press to filter under pressure, choose a deep layer filter plate with a pore size of no more than 0.9 μm (such as 0.6 μm), and control the temperature of the filter press to below -1°C. The FI precipitate was separated by filtration to obtain the FI supernatant;

[0040] Step 2. Separation of FⅡ+Ⅲ from th...

Embodiment 3

[0050] see figure 1 , the method for extracting human serum albumin with low-temperature ethanol, using human plasma as raw material, adopting low-temperature ethanol method and pressure filtration method to produce human serum albumin, the production operating pressure is less than 0.25Mpa, comprising the following steps:

[0051] Step 1. Isolation of FI from human plasma:

[0052] Control the temperature of the fused human plasma at 4.0°C, adjust the protein concentration to 65g / L, adjust the conductivity of the solution solution to 13.7ms / cm, adjust the pH value to 7.30, adjust the ethanol concentration to 10.0% by volume, and control the temperature to - 3.0°C, after the precipitation is complete, use a filter press to filter under pressure, choose a deep layer filter plate with a pore size of no more than 0.9 μm (such as 0.3 μm), control the temperature of the filter press to below -1°C, and filter Separate the FI precipitate to obtain the FI supernatant;

[0053] Step ...

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Abstract

The invention discloses a method for low-temperature extraction of human serum albumin employing ethanol. The method comprises the following steps: orderly separating out FI, FII+III and FIV from human plasma as a raw material, so as to obtain an FV sediment; dissolving the FV sediment, adjusting the protein content of the solution to 20-60g/L, the pH value to 4.50-4.70, and the ethanol concentration to 9.0-13.0; filtering by 3-5 stages, carrying out ultrafiltration dialysis on the solution and concentrating into required concentration, and then adding sodium caprylate, so that the sodium caprylate content in the solution is 0.140-0.180mmol/g of protein; adding sodium chloride, so that the osmotic pressure of the solution is 210-400 mOsmol/kg; adjusting the pH value of the solution to 6.40-7.40 and diluting into specified packaging concentration. By adopting the method disclosed by the invention, the extraction rate of the FV sediment is improved, and the yield of the human serum albumin is greatly improved.

Description

technical field [0001] The invention relates to a method for preparing human serum albumin using human plasma as a raw material, in particular to a method for extracting human serum albumin with low-temperature ethanol. Background technique [0002] The low-temperature ethanol method is one of the preparation methods of human plasma albumin. This method was invented by Professor Edwin J. Cohn of Harvard Medical School in the United States in 1940, so it is also called "Kong's method". The Kong method was originally used to separate bovine serum albumin and was subsequently applied to the separation of human plasma. The low-temperature ethanol method uses mixed plasma as the raw material to gradually reduce the acidity (from pH7.0 to pH4.0). Increase the concentration of ethanol (from 0 to 40%), while reducing the temperature (from 2°C to -2°C), various proteins are separated from the solution step by step in the form of components (crude products) under different conditions...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K1/14
CPCC07K14/765
Inventor 安康张宝献张其昌王猛滕世超郭心怡
Owner HUALAN BIOLOGICAL ENG CHONGQING
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