A method for preparing human serum albumin from urine

A technology for human serum albumin and urine protein, applied in the field of human serum albumin preparation, can solve problems such as application limitation, side reactions such as albumin allergy, unknown hidden dangers, etc. Effect

Active Publication Date: 2016-05-25
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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  • A method for preparing human serum albumin from urine
  • A method for preparing human serum albumin from urine
  • A method for preparing human serum albumin from urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] a Metal chelation affinity chromatography

[0045] Collect 100 kilograms of resin for urinary protein, rinse it with drinking water, and pack it into a column; eluting with 0.5M NaCl, collecting the elution peak, and measuring the albumin content of 5.60g.

[0046] Use an ultrafiltration membrane with a molecular weight cutoff of 30K for ultrafiltration and concentration. Adjust the pH of the ultrafiltration concentrate to 4.0 and conductance to 0.5mS / cm. The metal ions that have been balanced are Cu 2+ The metal chelating affinity chromatography column (ChelatingsepharoseFF); then rinse the metal chelating affinity chromatography column with the balance solution (equilibration solution formula: 0.2M acetate buffer, 2.0M NaCl, pH4.0), and collect the sample The penetrating fluid and the flushing penetrating fluid are washed with 0.2M phosphate buffer, 0.1M ammonium chloride (pH 5.0), and the metal chelate is eluted with 0.2M phosphate, 0.5M ammonium chloride (pH 6.0) Combine...

Embodiment 2

[0055] a Metal chelation affinity chromatography

[0056] Collect 100 kilograms of resin that adsorbs urine protein, rinse it with drinking water, and pack it into a column; eluting with 1.5M NaCl, collecting the elution peak, and measuring the albumin content in it is 4.8g.

[0057] Use an ultrafiltration membrane with a molecular weight cut-off of 30K for ultrafiltration and concentration. Adjust the pH of the ultrafiltration concentrate to 8.0 and conduction to 10mS / cm. The metal ions that have been balanced are Zn. 2+ The metal chelating affinity chromatography column (ChelatingsepharoseFF), and then using the balance solution (equilibration solution formula: 0.02M phosphate buffer, 0.2M NaCl, pH 8.0) to wash the metal chelating affinity chromatography column, collect the sample Permeate and rinse the penetrating fluid, rinse with 0.02M phosphate buffer, 0.5M ammonium chloride (pH 8.0), eluate metal chelation with 0.02M phosphate, 1.5M ammonium chloride (pH 8.4) Affinity chroma...

Embodiment 3

[0066] a Metal chelation affinity chromatography

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

[0068] Use an ultrafiltration membrane with a molecular weight cut-off of 30K for ultrafiltration and concentration. Adjust the pH of the ultrafiltration concentrate to 6.0 and conductance to 5.0mS / cm. The metal ions that have been balanced are Cu 2+ The metal chelating affinity chromatography column (ChelatingsepharoseFF); then rinse the metal chelating affinity chromatography column with the balance solution (equilibration solution formula: 0.1M phosphate buffer, 1.0M NaCl, pH 6.0), and collect the sample Permeate and rinse the penetrant, rinse with 0.1M phosphate buffer, 0.3M ammonium chloride (pH7.0), and elute metal chelate with 0.1M phosphate, 1.0M ammonium chloride (pH7.0) Affinity chromatography column, collect the elution ...

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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 manufacturing field of human albumin, in particular to a method for preparing human 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 largest protein drug used in the clinic. It is clinically used to treat diseases such as hypoalbuminemia, chronic nephritis, hepatitis, liver cirrhosis and advanced malignancies. In addition, human albumin is also used as an excipient and stabilizer , Immune adjuvant for vaccine production, components of serum-free cell culture medium, etc. [0003] Human albumin is mainly derived from plasma. According to market statistics in 2012, the global demand for human albumin is more than 600 tons per year. Except the United States, most other countries need to rely on partial ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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