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

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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 f

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  • Serum albumin degreasing method
  • Serum albumin degreasing method
  • Serum albumin degreasing method

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0019] Example 1

[0020] Test as a starting material as a starting material in the CoHN group IV.

[0021] The first step: Preparation of the Cohnium component IV supernatant: 1L plasma is separated from the classic CoHnium 6 method to separate to the component IV supernatant, co-acquisition component IV upper liquid about 3.15L, supernatant The ethanol volume concentration is 40%, the protein content is 10.0 g / L, pH of 6.0 ± 0.2, and the conductivity is about 3.34 ± 0.2 ms / cm.

[0022] Step 2: The solution of ethanol volume concentration of 40%, carbonamin is 10 mol / L, and the solution is adjusted to pH = 4.3 with acetic acid.

[0023] Step 3: Take 1.05L slowly from the solution of the second step to the first step in the first step IV supernatant, the ethanol volume concentration of ethanol in the final solution is 40%, the carbonamamamine concentration is 2.5 mol / L, and stirring, and The temperature is gradually reduced to -8 ° C, and the pH to 4.8 is adjusted with ace...

Example Embodiment

[0034] Example 2

[0035] The albumin precipitate prepared by low temperature ethanol is performed as a starting material:

[0036] The first step is to remove the blood albumin component V precipitate, dissolved with a solution of 40% aqueous ethanol solution, and the solution of sodium chloride is added to adjust the solution electrical conductivity, and the pH is adjusted to 6.0 ± 0.2 with acetic acid. The measured protein content was 19.8 g / l, and the conductivity was 3.0 ms / cm.

[0037] Step 2: The solution of ethanol volume concentration of 40%, carbonamin is 10 mol / L, and the solution is adjusted to pH = 4.3 with acetic acid.

[0038] In the third step: Take the solution of the second step 0.943L slowly added to the first step, the ethanol volume concentration of ethanol in the final solution is 40% concentration, carbonamamide 3 mol / L, continuously stir, and gradually reduce the temperature to -8 ° C or less, The solution pH = 4.8 ± 0.2 was adjusted with acetic aci...

Example Embodiment

[0049] Example 3

[0050] Test as a starting material with a weight concentration of 20% by weight of human serum albumin solution

[0051] The first step: 10% by weight of the human serum albumin solution 100 ml, the injection is diluted to 400 mL, and 95% ethanol is added to 290 ml to an ethanol volume concentration of 40%, and pH is adjusted to 6.0 ± 0.2 with acetic acid - acetate. About 29.8 g / l, the electrical conductivity is about 2.7 ms / cm, and about 672 ml of protein solution is obtained.

[0052] Step 2: The solution of ethanol volume concentration of 40%, carbonamin is 10 mol / L, and the solution is adjusted to pH = 4.3 with acetic acid.

[0053] In the third step: Take 168 ml of the second step to 168 ml of slowly added to the first step, the ethanol volume concentration of ethanol in the final solution is 40%, carbonamamine 2 mol / L, continuously stirred, and gradually reduce the temperature to -8 ° C or less, Acetic acid regulates pH to 4.8 ± 0.2.

[0054] Step ...

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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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