Improving the time integral of viable cell concentration by overcoming cell death, namely apoptosis, is one of the most widely used strategies for the efficient production of therapeutic proteins. By establishing stable cell lines that overexpress antiapoptotic genes or downregulate proapoptotic genes, the final product yields can be enhanced as cells become more resistant to environmental stresses. From the selection of high-expressing clones to verification of antiapoptotic activity, the method to construct a stable antiapoptotic cell line is discussed in this chapter.
Keywords: Apoptosis; Cell culture; Chinese hamster ovary cells; Overexpression; Selection; Suspension adaptation; Therapeutic proteins; Transfection; Western blot analysis; siRNA.
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