Staphylococcus protein A as well as purification preparation method and application thereof

A staphylococcal protein and protein technology, which is applied to the preparation method of peptides, biochemical equipment and methods, immunoglobulin, etc., can solve the problems of insufficient purity and IgG purity, and achieve simple preparation process, high purification efficiency, and durable The effect of high alkalinity
CN113512099AActive Publication Date: 2021-10-19赣江中药创新中心
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
赣江中药创新中心
Publication Date
2021-10-19

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Abstract

The invention discloses staphylococcus protein A as well as a purification preparation method and application thereof. The method comprises the following steps: connecting a protein Protein A gene modified by genetic engineering into an expression vector to construct a recombinant plasmid; introducing the recombinant plasmid into a prokaryotic host cell for expression and carrying out Ni affinity and ion-exchange chromatography purification, thereby obtaining the target protein with the purity greater than 95%. The purification preparation method has the characteristics of simple purification process, high yield and the like. Meanwhile, the prepared Protein A has good immunoglobulin IgG binding force, alkali resistance and thermal stability. And the affinity medium prepared by coupling the solid phase carrier with Protein A has the advantages of good IgG adsorption effect, high sample loading amount and alkali resistance. The Protein A protein prepared by the invention and a medium prepared by coupling the Protein A protein can play an important role in the field of monoclonal antibody purification and immunodiagnosis research.
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Description

technical field

[0001] The invention relates to the technical field of cloning and transforming Protein A in genetic engineering, in particular to a staphylococcal protein A, a purification preparation method and an application thereof. Background technique

[0002] In the middle of the 20th century, Jensen discovered a protein that could widely bind to human and rabbit serum antibodies. This protein was first called Protein A in 1964. Protein A is a cell wall protein of Staphylococcus aureus with a molecular weight of 42KDa. It can specifically bind to the Fc region of various immunoglobulins and weakly bind to the Fab region or light chain. Natural Protein A has 5 IgG binding domains E, D, A, B, C and a non-Fc binding domain of unknown function. The 5 different domains can bind the Fc fragment of IgG with strong specific affinity. A ProteinA molecule can bind at least two IgG molecules. At the same time, IgA, IgM or IgE may also combine with ProteinA. Protein A of this ...

Claims

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