Method for purifying antibody by denatured antigen affinity purification

Abstract

The invention relates to a novel method for purifying an antibody by denatured antigen affinity purification. In the antigen chromatographic column preparation process, the antigen is coupled with a solid-phase carrier in a urea-containing medium; and as the urea concentration gradually decreases, the antigen connected with the solid phase is utilized to implement in-situ renaturation, thereby ensuring that the subsequent antibody affinity purification process is completed in the water solution medium. The denatured antigen protein (such as prokaryotically-expressed inclusion body protein) can be directly coupled to the solid-phase carrier to purify the antibody, thereby simplifying the operation steps (the soluble antigen protein preparation process). The method has the advantages of high operability and low consumption. The obtained antibody has very high purity and specificity, and is suitable for laboratory preparation and industrial production.

Description

technical field [0001] The present invention is a technical scheme of antibody affinity purification with denatured antigen coupling solid-phase carrier reaction as the main innovation point, especially suitable for polyclonal antibody purification using insoluble protein prepared by prokaryotic expression as antigen, and belongs to antibody affinity purification technology field. Background technique [0002] There are many methods for antibody purification. In the early days, people used the salting out method, and later developed into the method of ion exchange chromatography. At present, the more commonly used method is to use Protein A (type A Staphylococcus aureus cell wall protein), G (type G gold Staphylococcus aureus cell wall protein), A / G or the antigen itself coupled to the activated matrix for antibody affinity purification. In comparison, antibodies obtained by antigen affinity purification have good affinity, strong specificity, and high purity, and are bette...

AI Technical Summary

Problems solved by technology

However, since most inclusion bodies are difficult to refold in aqueous solution, and antibody activity is easily damaged by denaturants, it has always been a difficulty to use denatured antigen protein (such as inclusion bodies) to affinity purify antibodies.

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