Abstract

The present invention relates to a method for obtaining high-purity and high-stability recombinant human albumin by adding medium- and long-chain fatty acid ligands and removing a protein having charge heterogeneity by means of anion or/and cation chromatography. The present invention is characterized in that a mixed solution is added to a sample collection solution, the mixed solution is a mixed solution of a fatty acid mixture and poloxamer; the molar ratio of oleic acid to myristic acid to sodium palmitate to sodium stearate to recombinant human albumin is 0.3:0.3:0.3:0.5:1; and truncated albumin and mismatched or modified albumin in recombinant albumin can be effectively removed by means of sequential use of ion exchange chromatography, so that a high-purity and high-stability human albumin sample is obtained.

IPC Classes  ?

• C07K 14/765 - Serum albumin, e.g. HSA
• C07K 1/18 - Ion-exchange chromatography
• C07K 1/14 - ExtractionSeparationPurification
• A61K 38/00 - Medicinal preparations containing peptides
• A61K 38/38 - Albumins
• A61P 7/00 - Drugs for disorders of the blood or the extracellular fluid
• C07K 14/76 - Albumins
• C12N 15/14 - Human serum albumins

Abstract

Provided is a method for improving the expression level of a recombinant human albumin, which comprises expressing the recombinant human albumin, and weakening, destroying or removing the function of at least one endogenous gene encoding a flocculating protein, and/or editing an endogenous gene or heterologous gene to overexpress superoxide dismutase; and/or editing an endogenous gene or heterologous gene to overexpress E3 ubiquitin ligase. Further provided are a corresponding cell and albumin.

IPC Classes  ?

• C07K 14/765 - Serum albumin, e.g. HSA
• C12N 15/81 - Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
• C12N 15/14 - Human serum albumins
• C12N 1/19 - YeastsCulture media therefor modified by introduction of foreign genetic material
• C12N 15/53 - Oxidoreductases (1)
• C12R 1/78 - Hansenula
• C12R 1/72 - Candida
• C12R 1/84 - Pichia

Abstract

The present invention provides a method for purification of a recombinant protein, in particular to a recombinant human albumin; the method includes: (a) adding aminoguanidine and a medium-long chain fatty acid to a sample containing the recombinant protein; and (b) chromatographing the obtained sample, where the chromatography is optionally performed with a chromatographic buffer solution containing aminoguanidine.

IPC Classes  ?

• C07K 1/16 - ExtractionSeparationPurification by chromatography
• C07K 14/765 - Serum albumin, e.g. HSA
• C07K 14/535 - Granulocyte CSFGranulocyte-macrophage CSF
• C07K 14/54 - Interleukins [IL]
• C07K 14/555 - Interferons [IFN]
• C07K 14/61 - Growth hormone [GH], i.e. somatotropin

Abstract

A preparation method of a human albumin-containing medical product comprises controlling a content of long-chain fatty acids in the medical product and adding poloxamer to the medical product. The long-chain fatty acid is one or more selected from myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid and arachidonic acid, and a molar ratio of the long-chain fatty acids and human albumin is 0.2-3.0. A content of the poloxamer is 10-500 μg/g of human albumin. The human albumin-containing medical product prepared by the method has improved stability and does not contain particle of over 30 nm in diameter is formed from aggregation and/or fibrillation of human albumin.

IPC Classes  ?

• A61K 38/38 - Albumins
• A61K 47/10 - AlcoholsPhenolsSalts thereof, e.g. glycerolPolyethylene glycols [PEG]PoloxamersPEG/POE alkyl ethers
• A61K 47/12 - Carboxylic acidsSalts or anhydrides thereof

Abstract

Disclosed is a method for purifying a recombinant protein, in particular recombinant human albumin. The method comprises: (a) adding aminoguanidine and a medium-long chain fatty acid to a sample containing a recombinant protein; and (b) performing chromatography on the resulting sample. The chromatography is optionally performed with a chromatography buffer containing aminoguanidine.

IPC Classes  ?

• C07K 14/765 - Serum albumin, e.g. HSA

Abstract

Disclosed is a method for purifying a recombinant protein, in particular recombinant human albumin. The method comprises: (a) adding aminoguanidine and a medium-long chain fatty acid to a sample containing a recombinant protein; and (b) performing chromatography on the resulting sample. The chromatography is optionally performed with a chromatography buffer containing aminoguanidine.

IPC Classes  ?

• C07K 14/765 - Serum albumin, e.g. HSA

Abstract

Provided is a method for improving the expression quality and reducing the degradation of recombinant human albumin. According to the method, the protein disulfide isomerase (PDI) is overloaded and expressed in a recombinant host cell, thereby helping to reduce incorrect shearing in a protein secretory pathway, reduce the degradation of proteins, and improve the expression quality of foreign proteins.

IPC Classes  ?

• C12N 1/19 - YeastsCulture media therefor modified by introduction of foreign genetic material
• C12N 15/81 - Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
• C12N 15/14 - Human serum albumins
• C12N 15/61 - Isomerases (5)
• C12R 1/84 - Pichia
• C12R 1/78 - Hansenula
• C12R 1/72 - Candida

Abstract

Disclosed is a method for preparing a preparation of human albumin, the method comprising controlling the content of a long chain fatty acid in the preparation; and adding a poloxamer. The long chain fatty acid is selected from one or more of myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid and arachidonic acid, and the molar ratio of the long chain fatty acid to human albumin is 0.2-3Ø The content of the poloxamer is 10-500 microgram per gram of the human albumin. The preparation of human albumin prepared by means of the method has an improved stability, and is free from a particle with a particle size of at least 30 nm formed by human albumin aggregation and/or fibrillation.

IPC Classes  ?

• A61K 38/38 - Albumins
• A61K 47/10 - AlcoholsPhenolsSalts thereof, e.g. glycerolPolyethylene glycols [PEG]PoloxamersPEG/POE alkyl ethers
• A61K 47/12 - Carboxylic acidsSalts or anhydrides thereof
• A61P 7/08 - Plasma substitutesPerfusion solutionsDialytics or haemodialyticsDrugs for electrolytic or acid-base disorders, e.g. hypovolemic shock

Abstract

Disclosed is a method for preparing a preparation of human albumin, the method comprising controlling the content of a long chain fatty acid in the preparation; and adding a poloxamer. The long chain fatty acid is selected from one or more of myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid and arachidonic acid, and the molar ratio of the long chain fatty acid to human albumin is 0.2-3.0. The content of the poloxamer is 10-500 microgram per gram of the human albumin. The preparation of human albumin prepared by means of the method has an improved stability, and is free from a particle with a particle size of at least 30 nm formed by human albumin aggregation and/or fibrillation.

IPC Classes  ?

• A61K 38/38 - Albumins
• A61K 47/10 - AlcoholsPhenolsSalts thereof, e.g. glycerolPolyethylene glycols [PEG]PoloxamersPEG/POE alkyl ethers
• A61K 47/12 - Carboxylic acidsSalts or anhydrides thereof
• A61P 7/08 - Plasma substitutesPerfusion solutionsDialytics or haemodialyticsDrugs for electrolytic or acid-base disorders, e.g. hypovolemic shock