Established The Platform Of Transient Transfection For Antibody Expressed In HEK293 PEAK Cells

Transfection is the process of deliberately introducing nucleic acids into cells (plasmid、DNA、mRNA、siRNA、microRNA), it is an important method of exogenous genetic materials expressing in cells. Transfection can be divided into transient transfection and stable transfection. In transient transfection, DNA is usually not integrated into the nuclear genome of host cell, the foreign DNA will be diluted through mitosis or degraded. Therefore, the exogenous gene can be expressed in host cells for obtaining available antibody samples in a short time. This is an important method in arly stage of antibody drug development. In stable transfection, DNA is usually integrated into the nuclear genome of host cell, With the cell genome replication of amplification, this is an important method for obtaining stable cell line expressing antibody.PEI, a cationic polymer was used as a new nonviral delivery vehicle for transfecting cells. It has many advantages, such as economic and utility, high transfection efficiency, low cytotoxicity, easy to operate, good repeatability. It does not influence the transfection efficiency with or without serum in medium. PEI is applicable to the most commonly used cell lines.Human embryonic kidney cells(HEK293 cells), has been widely used in cell biology research, the production of therapeutic proteins and gene therapy. HEK293 PEAK rapid cells are very easy to grow, and can growth in the environment without calcium ions. They are easily adapted in serum-free suspension culture, used as host cells for transient transfection expressing proteins.This research established engineering cell line for transient transfection for antibody by adapting with Gibco(?) FreeStyleTM 293 expression medium and limited dilution. The ratio of antibody plasmid and PEI(transfection reagent) was optimized to be that the ratio of 1:2(antibody plasmid:PEI) is the best result. The molecular weight of antibody expressed in HEK293 PEAK cells is about 150KD, and the biological activity was similar to control antibody(Herceptin). In summary we successfully established the platform for rapidly expressing antibody in HEK293 PEAK cells cultured in serum-free medium.