Method for producing recombinant protein A

A technology of recombinant protein and protein, which is applied in the field of high-density fermentation and protein separation and purification, to achieve the effects of low production cost, easy amplification, and high binding activity
CN101921818AActive Publication Date: 2010-12-22DALIAN UNIV OF TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
DALIAN UNIV OF TECH
Publication Date
2010-12-22

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Abstract

The invention relates to a method for producing recombinant protein A, which belongs to the technical field of biological engineering and provides a technological condition for fermenting and expressing the recombinant protein A in high density by using genetic engineering bacteria, and a method for separating and purifying the recombinant protein A in high purity by using Fc segment of IgG as anaffinity ligand by a one-step affinity chromatography method. By applying the method for producing the recombinant protein A, the culture density of the recombinant bacteria fermented in high densityreaches OD600nm=80-100; the dry weight of the bacteria is 40-50g / L; the expression quantity of the protein A occupies 30% to 50% of total protein of the bacteria; the quantity of the protein A produced by each liter of bacterial liquid reaches 3g; and the purity of the protein A detected by SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and HPLC (High Performance Liquid Chromatography) is more than 95%. The method has the advantages of large yield, low cost, high product quality, and the like, and provides a practicable approach for preparing the recombinant protein A.
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Description

technical field

[0001] The invention belongs to the technical field of bioengineering, and in particular relates to a high-density fermentation of genetically engineered bacteria and a method for separating and purifying proteins. Background technique

[0002] Staphylococcus aureus protein A (Staphylococcus aureus Protein A, SPA) is the cell wall binding protein of certain species of Staphylococcus aureus, which was studied by Danish scientist Klaus Jensen in the 1950s to study the cell wall structure of Staphylococcus aureus discovered when. Its basic structure is composed of the following three parts, namely signal peptide, antibody binding functional region and cell wall binding region. The antibody-binding functional region of protein A contains five homologous single domains E, D, A, B, and C, and each single domain can act independently with immunoglobulin G (IgG). The binding of the protein to IgG is mainly based on the specific binding ability to the Fc region of I...

Claims

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