Recombinant Protein A protein and preparation method of affinity chromatography medium

A chromatographic medium and protein technology, which is applied in the field of protein engineering, can solve the problems of reduced lifespan, unsatisfactory stability, and destruction of the advanced structure of Protein A, and achieve excellent antibody binding ability, good chemical stability, and improved chemical stability.
CN113278052AActive Publication Date: 2021-08-20PINGHU YOUPU BIOTECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
PINGHU YOUPU BIOTECH CO LTD
Publication Date
2021-08-20

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Abstract

The invention discloses a recombinant Protein A protein and a preparation method of an affinity chromatography medium. The protein is a mutant of parent immune globulin-binding protein as defined by SEQ ID No 1 and SEQ ID No 2; and the preparation method of the affinity chromatography medium comprises the following steps: S1, synthesizing genes of the recombinant Protein A protein, S2, respectively constructing expression vectors by using the genes synthesized in the S1, S3, culturing, expressing and purifying to obtain a recombinant Protein A solution with a corresponding sequence of SEQ ID, and S4, preparing the affinity chromatography medium by using the recombinant Protein A. The preparation method has the advantages that the chemical stability of the Protein A in alkali liquor is greatly improved by redesigning an amino acid sequence, so that the Protein A with a specific binding effect on an antibody is provided, the Protein A has good chemical stability under an alkaline condition, the binding load of the recombinant Protein A and affinity chromatography medium is remarkably improved, in-place cleaning under 0.5-1.0 M NaOH can be tolerated, and the IgG binding load is 60-90 mg / ml.
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Description

technical field

[0001] The invention relates to the technical field of protein engineering, in particular to a method for preparing recombinant Protein A protein and an affinity chromatography medium. Background technique

[0002] Antibody drugs have excellent curative effect and safety on diseases, and have gradually become important in the treatment of tumors, cardiovascular diseases, especially autoimmune diseases, and are currently the class of drugs with the highest growth rate among biological drugs . Thanks to the construction of high-expression cell lines, improvement of culture medium, and continuous expansion of reactor scale, monoclonal antibody drug technology has developed rapidly in the past few decades. In the preparation process of monoclonal antibody drugs, the purification process of antibodies is particularly important, and the capture of antibodies plays a key role. The Protein A affinity chromatography medium is currently the most widely used and most ...

Claims

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