AbstractAs a rapidly developing cell engineering technique, cell electrofusion has been increasingly applied in the field of hybridoma preparation in recent years. However, it is difficult to completely replace the polyethylene glycol‐mediated cell fusion using electrofusion due to the high operation requirements, high cost of electrofusion instruments, and lack of prior reference research work. The key elements limiting electrofusion in the field of hybridoma preparation also introduce practical complications, such as the use/choice of electrofusion instruments, setup/optimization of electrical parameters, and precise control of cells. This review summarizes the state of the art of cell electrofusion in hybridoma preparation based on recent published literature, mainly focusing on electrofusion instruments and their components, process control and characterization, and cell treatment. It also provides new information and insightful commentary critically important for further electrofusion development in the field of hybridoma preparation.

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