Efficient electrotransfection method for HEK293F suspension cells

Abstract

The invention provides an efficient electrotransfection method for HEK293F suspension cells. According to the method, the voltage intensity, the number of pulses, the pulse interval duration, the electric shock duration and the like during electrotransfection are optimized, so that efficient electrotransfection of the HEK293F cells subjected to suspension culture is achieved; and without preparing an electrotransfection buffer solution during electrotransfection, the relatively high transfection efficiency can be obtained. The method can be used for different plasmid transfected and suspension cultured HEK293F cells, and eukaryotic expression of a large number of target proteins (such as protein products of antibodies, enzymes and the like) is obtained.

Description

technical field [0001] The invention belongs to the field of transgenic technology, in particular to a high-efficiency electrotransfection method for HEK293F suspension cells. Background technique [0002] The liposome transfection method is currently the most used transfection method. Liposomes encapsulate DNA to form DNA / liposome complexes, and cells transfer DNA into cells by endocytosis. However, the quality of DNA has a great influence on the transfection efficiency. Small doses of endotoxin, salt ions, proteins and polysaccharides will affect the formation of complexes. Therefore, long-term contact of low-quality DNA with cells will lead to increased cell death. Reduced transfection efficiency. Moreover, the price of liposomes on the market is relatively high, which is not suitable for large-volume transfection. Although PEI is cheap, its transfection efficiency is low and its cytotoxicity is very high. Calcium phosphate transfection of cells requires large amounts ...

AI Technical Summary

Problems solved by technology

Dendritic cells are the most powerful antigen-presenting cells found so far, and HepG2 cells are an important cell model for liver cancer research. Both are adherent cells and are the focus of tumor biotherapy; however, targeting dendritic cells and HepG2 When cells are electrotransfected, due to their high cell density, it is difficult for some cells to reach the threshold

Under the reported optimal electroporation conditions for DC cells and HepG2 cells, the electroporation efficiency is 50%-60%, but there are problems such as the need for electroporation buffer during electroporation, incubation at 4°C before electroporation, or large amount of nucleic acid.

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