Inclusion body renaturation method and kit

Abstract

The invention discloses an inclusion body renaturation method and a kit. The method comprises the following steps: inducing a target protein to express in a host cell, and then splitting the host cell to obtain the target protein, dissolving the target protein; purifying the target protein by nickel column chromatography, and carrying out gradient elution by using an elution buffer solution containing sodium dodecyl sarcosinate and imidazole; and collecting the eluent containing the target protein, filling the eluent into a dialysis bag, and putting a renaturation buffer solution containing Tris-HCl and EDTA into the dialysis bag to dialyze and renaturate the target protein to obtain the renatured protein. Aiming at the problems that an existing inclusion body renaturation method is complex in operation, not easy to control, high in cost, low in yield, poor in activity and the like, the elution buffer solution and the renaturation buffer solution are optimized to play a synergistic effect, the renaturation effect of the inclusion body is effectively improved, the purity and activity of target protein are improved, and the method is simple to operate, and low in cost, and has a relatively high application value.

Description

technical field [0001] The invention belongs to the technical field of protein engineering, and in particular relates to a renaturation method and a kit for inclusion bodies. Background technique [0002] The E. coli protein expression system has a clear gene background, easy cultivation and control, simple transformation operation, high expression level, low cost, short cycle time, and high expression efficiency. It has become an important tool for researchers to study recombinant proteins. However, when exogenous proteins are highly expressed in E. coli, they often form insoluble and biologically inactive inclusion bodies. Inclusion bodies must go through the process of denaturation and renaturation in order to obtain proteins with correct structure and activity. At present, the widely used refolding methods include dilution refolding, dialysis refolding and on-column refolding. Dilution refolding is to quickly dilute and dissolve inclusion body protein with buffer to red...

AI Technical Summary

Problems solved by technology

[0003] The present invention aims at technical problems such as complicated operation and difficult control, high cost, low yield of obtained protein, and poor activity in the existing inclusion body renaturation method, and provides a renaturation method of inclusion body, through the elution buffer Optimize the selection of the components of the refolding buffer, replace the high concentration of urea in the traditional elution buffer with low concentration of sodium lauryl sarcosine, and omit the traditional concentration gradient in the refolding buffer The changed urea can effectively improve the renaturation effect of the inclusion body, improve the purity and activity of the target protein, and the method is simple to operate, low in cost, and has high application value

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