A kind of renaturation solution for improving the renaturation rate of prokaryotic inclusion body expression and its preparation method and application

Abstract

The invention belongs to the technical field of biology, and particularly relates to a renaturation solution capable of enhancing protocaryon inclusion body expression renaturation rate, and a preparation method and application thereof. The invention provides an optimized renaturation solution. The renaturation solution can nearly double the semi-inhibition rate of the crude product, and obviously enhances the preparation efficiency and yield returns of the recombinant protein products for the research units and production enterprises. The preparation method of the renaturation solution is simple, and is suitable for demands of large-scale industrial production. When the renaturation solution and renaturation method are used for producing HER2-positive mammary-cancer-targeted drugs, the problem of low renaturation efficiency in TP25 inclusion body expression is effectively solved, and the recovery rate of active proteins in the renaturation solution is enhanced.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a renaturation solution for improving the renaturation rate of prokaryotic inclusion body expression, a preparation method and application thereof. Background technique [0002] Prokaryotic expression is a common heterologous expression method. Using this expression system can achieve the purpose of fast, large-scale and high-efficiency acquisition of recombinant proteins, and has the advantages of short time-consuming and strong controllability, so many companies choose this expression system. , but this expression method is very easy to form inclusion bodies during the expression process. Although inclusion bodies indicate a large amount of expression, the activity recovery rate after denaturation is greatly reduced, which affects production efficiency and income, and increases The production cost of the product. [0003] There are more than 200,000 new cases of breast cancer ever...

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Problems solved by technology

[0002] Prokaryotic expression is a commonly used heterologous expression method. Using this expression system can achieve the purpose of obtaining recombinant proteins quickly, in large quantities, and efficiently, and has the advantages of short time-consuming and strong controllability, so many companies choose this expression system. , but this expression method is very easy to form inclusion bodies during the expression process. Although inclusion bodies indicate a large expression level, the recovery rate of activity after denaturation is greatly reduced, which affects production efficiency and income, and increases Product production cost

[0003] There are more than 200,000 new cases of breast cancer every year in my country, 30% of which are HER2-positive. TP25 is a new targeted anti-tumor drug for HER2-positive breast cancer. It is a class of drugs obtained through prokaryotic expression using modern genetic engineering techniques. However, because inclusion bodies are easily formed during the expression process, the activity recovery rate is greatly reduced, which affects the production efficiency and income, and increases the production cost of the product. Therefore, it is necessary to find an effective preparation of HER2-positive breast cancer targeting The renaturation solution and renaturation method of the drug TP25 inclusion body can not only improve the protein renaturation rate but also effectively reduce the renaturation cost, which has very important practical significance in production

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