Preparation method of recombinant human C-reactive protein

Abstract

The invention discloses a preparation method of recombinant human C-reactive protein. The method comprises the following steps: acquiring a protein sequence of the human C-reactive protein from NCBI;artificially optimizing and synthesizing a C-reactive protein encoding gene according to codon bias characteristics of escherichia coli; further constructing an expression vector of the human C-reactive protein; converting the expression vector into the escherichia coli for induced expression; and purifying an expression product to obtain pure recombinant protein. ELISA detection results show thatthe immunological characteristics of the expressed and purified recombinant human C-reactive protein are very similar to those of natural human C-reactive protein, and the detection requirements canbe met. In addition, the purification process of the recombinant human C-reactive protein is beneficial to quality control and the requirement of industrial production are met.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing recombinant human C-reactive protein. Background technique [0002] C-reactive protein (C-reactive protein, CRP) refers to some proteins (acute proteins) that rise sharply in plasma when the body is infected or tissue damaged. C-reactive protein can activate complement and strengthen the phagocytosis of phagocytic cells to play an opsonizing role, thereby removing pathogenic microorganisms invading the body and damaged, necrotic, and apoptotic tissue cells, and playing an important protective role in the body's natural immune process. [0003] At present, C-reactive protein is mainly extracted directly from human blood. Since the content of C-reactive protein in normal human serum is extremely low, below 10ug / ml, and human blood is inconvenient to obtain, it cannot meet the needs of a large number of clinical and scientific research. In the prior art, there ar...

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Problems solved by technology

[0003] At present, C-reactive protein is mainly extracted directly from human blood. Since the content of C-reactive protein in normal human serum is extremely low, below 10ug / ml, and human blood is inconvenient to obtain, it cannot meet the needs of a large number of clinical and scientific research.

In the prior art, there are also reports of expressing C-reactive protein through prokaryotic gene recombination, but because C-reactive protein exists in the form of inclusion bodies, it is difficult to renature during the protein purification process, and most of them are in the form of multimers after renaturation Exist, does not have the immunogenicity of natural CRP, cannot specifically bind to CRP antibody, cannot be tested by ELISA, and cannot be used for clinical testing

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