Recombination protein A gene and preparation of expression product thereof

A recombinant protein and gene expression technology, applied in the field of bioengineering, can solve the problems of little influence on the activity of protein A ligand, great harm to the human body and the environment, and low activation efficiency, and achieve strong binding specificity, short expression time, and high purity and high yield
CN101760466AActive Publication Date: 2010-06-30AGTC GENE TECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
AGTC GENE TECH CO LTD
Publication Date
2010-06-30

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Abstract

The invention relates to the technical field of biological engineering. The B structure domain gene monomer of protein A is optimized according to the preference of a colon bacillus colon, six series bodies are constructed and inserted into a thermally induced colon bacillus vector pBV220, and a colon bacillus expression vector containing the gene and a converted colon bacillus DH5alpha recombination strain thereof are constructed. And a method for producing the recombination protein A using the strain is also provided. The total soluble bacterial protein amount of the soluble recombination protein A produced by the strain can be more than 80 percent, and the recombination protein A can be purified to over 95% just by subsequent nickel ion chelate chromatography and molecular sieve column chromatography. Moreover, the protein has the advantages of high expression amount, low cost, easy purification and the like. A recombination protein A affinity stuffing prepared by using the protein has the advantages of high human IgG absorption capacity, low proA dropping and the like. The invention provides a practical and feasible approach to obtain the recombination protein A with high quality and low price as an antibody (medicine) purifying medium.
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Description

Technical field:

[0001] The invention belongs to the technical field of bioengineering, and relates to a recombinant protein A gene, a vector containing the recombinant gene, a transformed bacterial strain and a product expressed by the recombinant protein A gene, a purification method thereof, and preparation and use of an affinity filler. Background technique:

[0002] Staphylococcal Protein A (Staphylococcal Protein A, SPA) is a protein isolated from the cell wall of Staphylococcus aureus. In 1940, Vevwey found that some Staphylococcus aureus contained a substance that could form a precipitate with normal human serum in a two-way diffusion test. Jensen also discovered a similar phenomenon in 1959 and named it the A antigen. In 1963, Lofkvist and others isolated substance A, proving that it is a protein and different from sugar; in 1960, Grov named it staphylococcal protein A, referred to as SPA (ProteinA). The gene encoding SPA was cloned in 1983 and expressed in E. col...

Claims

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