Preparation method of SLAMF6 protein

Abstract

The invention discloses a preparation method of SLAMF6 protein expressed through eukaryocyte secretion. The preparation method comprises the steps that 1, the SLAMF6 protein is subjected to sequence optimization; 2, a cell strain for expressing the SLAMF6 protein is obtained through eukaryon resistance screening; 3, the cell strain is cultured, and the cell strain secreta SLAMF6 protein is obtained. The defects that prokaryotic expression is adopted in a traditional SLAMF6 protein production method, expression is poor, renaturation is difficult, and protein obtained through purification lacks glycosylation and cannot meet demands are overcome. According to the preparation method, a mammal expression system is adopted, through vector reconstruction, the cell strain for secretory expression of the protein is screened out of HEK293 cells, the problem that SLAMF6 protein is poor in expression is solved, and the problem of difficult renaturation is solved through secretory expression; meanwhile, recombinant protein of humans is expressed with cells of humans, more accurate glycosylation is achieved, and the research and development demand is met.

Description

technical field [0001] The invention relates to a recombinant protein, in particular to a preparation method of the SLAMF6 protein. Background technique [0002] SLAMF6 is a single-chain membrane protein belonging to the CD2 subfamily of the immunoglobulin superfamily. SLAMF6 is expressed in all NK cells, T cells and B lymphocytes. SLAMF6 is a homodimer, and SLAMF6 mediates the phosphorylation process of tyrosine together with SH2D1A and SHPs. As a co-receptor, SLAMF6 plays an important role in the activation process of NK cells. SLAMF6 also mediates the inhibitory response in NK cells derived from lymphoproliferative patients, so it is very important to express the protein through recombinant expression and study the function of the protein in vitro . [0003] According to the host cell type, gene expression systems can be roughly divided into prokaryotic, yeast, plant, insect and mammalian cell expression systems. Usually, prokaryotic expression systems are used to expr...

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Problems solved by technology

However, in most cases of prokaryotic expression of membrane proteins, the expressed protein is in the form of inclusion body, and renaturation is difficult, and the protein expressed by the prokaryotic system has no glycosylation process, so it is difficult to produce recombinant protein with biological functions , the traditional method of producing SLAMF6 protein is prokaryotic expression, poor expression, difficult renaturation, and the purified protein lacks glycosylation, which cannot meet the demand

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