Solubilization and purification of HBx protein core segment in HBV and monoclonal antibody

Abstract

The invention belongs to the field of a biological technology, and particularly relates to a purification method of an HBx protein core segment HBx delta N delta C in HBV and an HBx delta N delta C monoclonal antibody. A formula and purification process of various buffered solutions in an original purification method are changed, so that HBx delta N delta C samples being active, stable, uniform, high in purity and solitary are obtained, and standard large-scale production and preparation are convenient. The obtained HBx delta N delta C are used for further preparing a mouse sourced HBx delta Ndelta C monoclonal antibody and hybridoma strains which can stably express the mouse sourced HBx delta N delta C monoclonal antibody. The HBx delta Ndelta C and the HBx delta N delta C monoclonal antibody can be used as a scientific research reagent, the mouse sourced HBx delta N delta C monoclonal antibody can also be used as a detection reagent of hepatitis b virus HBV infection diseases, and the mouse sourced HBx delta N delta C monoclonal antibody is humanized, so that humanized HBx delta N delta C monoclonal antibody can be obtained and can be used as a scientific research reagent, a detection reagent for HBV infection diseases and an auxiliary treatment preparation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for solubilization and purification of the HBx protein core segment HBxΔNΔC in hepatitis B virus (HBV), and an HBxΔNΔC monoclonal antibody. Background technique [0002] The renaturation method of inclusion body protein is a traditional method for solubilizing inclusion body protein. This method first uses high concentration denaturant to solubilize the inclusion body protein, for example, use a buffer containing 8 M urea or 6 M thiourea to dissolve the inclusion body protein. Afterwards, through the process of removing the high-concentration denaturant, the inclusion body protein is refolded from a denatured state to a soluble state, such as using buffer dialysis without a denaturant to remove the denaturant, so that the inclusion body protein is refolded soluble. Due to the lack of understanding of the process and rules of protein folding and renaturation, th...

AI Technical Summary

Problems solved by technology

However, the protein samples obtained by combining these fusion proteins with HBx have the following disadvantages: the concentration usually can only reach about 1 mg / ml, HBx will precipitate immediately after the fusion protein is cut off by the enzyme, and larger fusion proteins may affect HBx and other proteins , nucleic acid interactions

[0006] A method for solubilizing and purifying HBxΔNΔC is reported in the published literature. The HBxΔNΔC obtained by this method is highly soluble in the solution, but the obtained HBxΔNΔC sample is unstable, easily degraded, inhomogeneous, not high enough in purity, and not functional. Stablize( figure 1 ), it is impossible to produce and prepare active, stable, high-uniformity, and high-purity protein preparations on a large scale. It is only suitable for small-scale laboratory research and has no commercial application prospects.

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