Serum albumin degreasing method

A serum albumin and albumin technology, applied in the field of serum albumin purification, can solve the problems of cumbersome operation and low processing efficiency

Pending Publication Date: 2022-01-28
广东卫伦生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] CN112759643A discloses a method for degreasing serum albumin. Under the condition of surfactant and / or organic solvent solution, serum albumin is separated from the conjugate by tangential flow filtration technology, so that the fatty acid content in albumin can be reached Not higher than the level of 5 μg / g albumin, but the program is cumbersome to operate and the processing efficiency is low

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  • Serum albumin degreasing method
  • Serum albumin degreasing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The test was carried out with Cohn's Fraction IV supernatant as starting material:

[0021] Step 1: Preparation of Cohn's Fraction IV supernatant: 1L of plasma was separated by low-temperature ethanol into Fraction IV supernatant by the classic Cohn's 6 method, and a total of about 3.15 L of Fraction IV supernatant was obtained. The volume concentration of ethanol is 40%, the protein content is 10.0g / L, the pH value is 6.0±0.2, and the conductivity is about 3.34±0.2 mS / cm.

[0022] Step 2: Prepare a solution of 40% ethanol volume concentration and 10 mol / L carbonamide, and adjust the solution to pH=4.3 with acetic acid.

[0023] The third step: Take 1.05L of the solution in the second step and slowly add it to the supernatant of component IV in the first step. The volume concentration of ethanol in the final solution is 40%, and the concentration of carbonamide is 2.5mol / L. Stir constantly, and Gradually lower the temperature to below -8°C, and adjust the pH to 4.8 wit...

Embodiment 2

[0035] The albumin precipitate prepared by the low-temperature ethanol method was used as the starting material for the degreasing test:

[0036] Step 1: Take about 200g of the precipitate of human serum albumin component V, dissolve it with 2.2L of ethanol aqueous solution with a volume concentration of 40%, add 2.667g of sodium chloride to adjust the conductivity of the solution, and adjust the pH to 6.0±0.2 with acetic acid , the measured protein content was 19.8g / L, and the conductivity was 3.0mS / cm.

[0037] Step 2: Prepare a solution of 40% ethanol volume concentration and 10 mol / L carbonamide, and adjust the solution to pH=4.3 with acetic acid.

[0038] Step 3: Take 0.943L of the solution in the second step and slowly add it to the solution in the first step. The volume concentration of ethanol in the final solution is 40%, and carbonamide is 3mol / L. Stir continuously, and gradually reduce the temperature to below -8°C. Adjust the pH of the solution to 4.8±0.2 with ace...

Embodiment 3

[0050] The human serum albumin solution with a weight concentration of 20% was used as the starting material for the test

[0051] Step 1: Take 100ml of human serum albumin solution with a weight concentration of 20%, dilute it to 400ml with water for injection, add 290ml of 95% ethanol until the volume concentration of ethanol is 40%, adjust the pH to 6.0±0.2 with acetic acid-sodium acetate, and measure the protein content It is about 29.8g / L, the conductivity is about 2.7mS / cm, and about 672ml of protein solution is obtained.

[0052] Step 2: Prepare a solution of 40% ethanol volume concentration and 10 mol / L carbonamide, and adjust the solution to pH=4.3 with acetic acid.

[0053] Step 3: Take 168ml of the solution of the second step and slowly add it to the solution of the first step. The volume concentration of ethanol in the final solution is 40%, and carbonamide is 2mol / L. Stir continuously, and gradually reduce the temperature to below -8°C. Acetic acid was used to ad...

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Abstract

The invention provides a simple and rapid serum albumin degreasing method. The method comprises the following steps: adding ethanol and carbamide into Cohn component IV supernate, albumin precipitate or albumin dissolving solution to prepare a solution with ethanol volume concentration of 30-50% and carbamide concentration of 1.0-3.0 mol / L, adjusting the pH value of the solution to 4.8 + / - 0.2 with acetic acid, cooling to-10 to -8 DEG C to precipitate albumin, filtering to obtain precipitate, washing and degreasing serum albumin by using a washing solution with ethanol volume concentration being 30-50%, the sodium chloride concentration being 100-150 mmol / L and the isopropanol concentration being 0.5-1mol / L and the pH value adjusted to 4.8 + / -0.2 by using acetic acid, dissolving and renaturating, and carrying out ultrafiltration dialysis to obtain degreased serum albumin of which the fatty acid content is reduced to 0.005 mol / mol.

Description

technical field [0001] The invention relates to a serum albumin purification method, especially a serum albumin degreasing method. Background technique [0002] Serum albumin is synthesized by the liver of the human body. It is the protein with the highest content in plasma, accounting for about 50-60% of the total plasma protein content. Physiological functions such as blood coagulation function and maintenance of capillary permeability. [0003] In blood and interstitial fluid, due to the unique structure of serum albumin molecules, it can bind many substances, including fatty acids, amino acids, Ga 2+ , Mg 2+ , steroids and their derivatives and other endogenous substances, as well as antibiotics, anticoagulants, heart drugs and other drugs. However, the binding site and binding capacity of serum protein molecules are limited, so the ligand molecules that bind to the same site may compete with each other or replace each other. [0004] In human serum albumin, medium a...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K1/36C07K1/34C07K1/30
CPCC07K14/765Y02A50/30
Inventor 黄锦程陈成坤管红玲李晓芝王文杰黄光义
Owner 广东卫伦生物制药有限公司
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