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Method for preparing human albumin from urine

A technology of human serum albumin and urinary protein, applied in the field of preparation of human serum albumin, can solve problems such as application limitations, side effects such as albumin allergy, unknown hidden dangers, etc., and achieve improved stability, reduced cost, and convenient operation Effect

Active Publication Date: 2014-04-02
YANGZHOU AIDEA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the genetic engineering products of albumin have made great progress. EP0612761, EP0570916, US6638740B1, etc. have all disclosed methods for purifying recombinant human serum albumin, but the albumin produced by these methods is prone to residual traces of heterologous proteins, causing allergies, etc. Side effects, the presence of recombinant albumin's uncertain physiological function can easily cause unknowable hidden dangers, so its application is severely limited

Method used

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  • Method for preparing human albumin from urine
  • Method for preparing human albumin from urine
  • Method for preparing human albumin from urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] aMetal chelate affinity chromatography

[0045] Collect 100 kg of resin that absorbs urinary protein, rinse it with drinking water, and pack it into a column; use 0.5 M NaCl for elution, collect the elution peak, and measure the albumin content in it to be 5.60 g.

[0046] Use an ultrafiltration membrane with a molecular weight cut-off of 30K for ultrafiltration concentration, adjust the pH of the ultrafiltration concentrate to 4.0, and the conductivity to 0.5mS / cm. The metal ion that has been balanced on the surface is Cu 2+ The metal chelating affinity chromatography column (Chelating sepharose FF); then use the equilibrium solution (equilibrium solution formula: 0.2M acetate buffer, 2.0MNaCl, pH4.0) to wash the metal chelating affinity chromatography column, collect Load the breakthrough and wash the breakthrough, rinse with 0.2M phosphate buffer, 0.1M ammonium chloride (pH5.0), and elute with 0.2M phosphate, 0.5M ammonium chloride (pH6.0) Metal chelate affinity chr...

Embodiment 2

[0055] aMetal chelate affinity chromatography

[0056] Collect 100 kg of resin that absorbs urinary protein, rinse it with drinking water, and pack it into a column; use 1.5M NaCl to elute, collect the elution peak, and measure the albumin content in it to be 4.8g.

[0057] Use an ultrafiltration membrane with a molecular weight cutoff of 30K for ultrafiltration concentration, adjust the pH of the ultrafiltration concentrate to 8.0, and the conductivity to 10mS / cm, and the metal ion that has been balanced on the surface is Zn 2+ The metal chelating affinity chromatography column (Chelating sepharose FF), and then use the equilibrium solution (equilibrium solution formula: 0.02 M phosphate buffer, 0.2M NaCl, pH8.0) to wash the metal chelating affinity chromatography column, and collect the Sample breakthrough and wash penetration, wash with 0.02M phosphate buffer, 0.5M ammonium chloride (pH8.0), and elute metal with 0.02M phosphate, 1.5M ammonium chloride (pH8.4) Chelate affin...

Embodiment 3

[0066] aMetal chelate affinity chromatography

[0067] Collect 5 tons of resin that absorbs urinary protein, rinse it with drinking water, and pack it into a column; use 0.8 M NaCl for elution, collect the elution peak, and measure the albumin content in it to be 216.00g.

[0068] Use an ultrafiltration membrane with a molecular weight cut-off of 30K for ultrafiltration concentration, adjust the pH of the ultrafiltration concentrate to 6.0, and the conductivity to 5.0mS / cm. The metal ion that has been balanced on the surface is Cu 2+ The metal chelating affinity chromatography column (Chelating sepharose FF); then use the balance solution (equilibrium solution formula: 0.1M phosphate buffer, 1.0M NaCl, pH6.0) to wash the metal chelation affinity chromatography column, and collect the Sample breakthrough and wash penetration, wash with 0.1M phosphate buffer, 0.3M ammonium chloride (pH7.0), and elute metal with 0.1M phosphate, 1.0M ammonium chloride (pH7.0) Chelate affinity chr...

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Abstract

The invention discloses a method for preparing human albumin from urine. Resins absorbing urine protein are collected for cleaning and desorbing; elution peaks are collected for ultrafiltration and concentration; the pH is adjusted; the electric conductivity is implemented; the human albumin is prepared by the metal chelating affinity chromatography, the hydrophobic chromatography and the anion exchange chromatography. The method has the advantages of low cost, high yield, no immunogenicity and stable quality, is suitable for large-scale production operation, and develops a brand new channel for the source of the albumin.

Description

technical field [0001] The invention relates to the field of production of human serum albumin, in particular to a method for preparing human serum albumin from urine. Background technique [0002] Human Serum Albumin (HSA) is the most important functional protein in human blood. Its main physiological function is to maintain blood osmotic pressure, combine and transport small molecule nutrients and drugs. HSA is currently the most widely used protein drug. It is clinically used to treat diseases such as hypoalbuminemia, chronic nephritis, hepatitis, liver cirrhosis and advanced malignant tumors. In addition, human albumin is also used as an excipient and stabilizer , immune adjuvants for vaccine production, components of serum-free cell culture medium, etc. [0003] Human serum albumin is mainly derived from plasma. According to market statistics in 2012, the global demand for human serum albumin is more than 600 tons per year. Except for the United States, most other coun...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K1/22C07K1/20C07K1/18
CPCC07K14/765
Inventor 苗丕渠侯晓彦池正昌沈小宁
Owner YANGZHOU AIDEA BIOTECH
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