KMD Bioscience Co., Ltd.
400-621-6806
2023-05-26 Hits(347)
Mammalian cells are systems that express human genes. The advantage of mammalian cells is to promote the correct folding of protein and provide accurate post-translational modifications, such as glycosylation and acetylation. It can secrete mature protein out of cells by constructing signal peptide, and easily obtain high-activity protein by affinity purification. Recombinant proteins based on mammalian cell expression system are widely used in the production of therapeutic recombinant proteins, because their advanced structure, post-translational modification, physical and chemical properties and biochemical activities are more similar to those of natural higher biological proteins. According to the temporal and spatial differences of target protein expression, mammalian cell expression system can be divided into transient expression system, stable expression system and induced expression system.
Because the cells expressed by mammalian cells need to grow steadily and continuously, and the expression of target genes can be realized by introducing foreign genes, it is necessary to obtain suitable cells by screening and optimizing animal tissues. Commonly used cell lines include HEK293 (human embryonic kidney cell) and CHO (Chinese hamster ovary cell), both of which are cultured in suspension cells and can produce several grams of protein in one month. However, mammalian expression system also has some shortcomings, such as high experimental cost, long expression cycle and high technical difficulty. The main problems of expressing recombinant protein in mammalian cells are low transfection efficiency and low protein level. However, with the development of scientific research, HEK293 cells and CHO cells have been developed into effective transient and stable expression systems respectively.
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Means of Expression |
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Transient Expression |
HEK293 cell, also called human embryonic kidney cell 293, is a cell line derived from human embryonic kidney cell. Serum-free suspension culture was used.
CHO-S cells, which originated from chinese hamster ovary cell, were domesticated into serum-free suspension culture.
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Stable Expression |
CHO-S and CHO-K1 are derived from CHO. The cell line has simple culture conditions, moderate adhesion strength and easy transfection.
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Transient transfection uses liposome, calcium phosphate or PEI as transfection reagent. HEK293 cells are relatively simple, have the advantages of higher growth density and faster growth rate, are easy to culture and transfect, and are widely used in the production of protein, vaccines and anticancer agents, but it is difficult to expand the scale. CHO cells belong to fibroblasts, are non-secretory cells, and rarely secrete endogenous proteins, which is conducive to the separation and purification of target proteins, and can be used to stably prepare a large number of recombinant proteins. At present, CHO cells are one of the most successful cells expressing exogenous proteins.
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