| Abstract | Transfection is a process of introducing nucleic acid into eukaryotic cells using various chemical or physical methods. As a nonviral delivery system, transfection has gained popularity. The cost-effective and rapid production of recombinant human therapeutic proteins in sufficient quantities has been one of the challenges faced in biopharmaceutical industries. Transient cell transfection met the challenge by achieving these results that avoids a lengthy and costly procedure of developing stable transfectants. To date, transfection technology has also been applied to produce r-proteins, antibodies, and viral vectors for therapeutic, gene therapy, and vaccine applications. This article outlines classical transfection methods and details two most popular methods: calcium phosphate precipitation method and polyethylenimine (PEI)–DNA condensation method. Both methods are applicable for the production of r-proteins by using anchorage-dependent as well as suspension-growing cell lines. However, the PEI-based method has been preferred among the researchers to further the advancement of the transfection technology and produce r-proteins in streamlined large-scale serum-free suspension cultures. Recent advances in large-scale transfection technology are reviewed and optimization techniques are mentioned that include selection of cell line, plasmid preparation, and choice of medium and transfection reagent. |
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