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Molecular engineering of exocytic vesicle traffic enhances the productivity of Chinese hamster ovary cells

A complex vesicle trafficking system manages the precise and regulated distribution of proteins, membranes and other molecular cargo between cellular compartments as well as the secretion of (heterologous) proteins in mammalian cells. Sec1/Munc18 (SM) proteins are key components of the system by regulating membrane fusion. However, it is not clear how SM proteins contribute to the overall exocytosis. Here, functional analysis of the SM protein Sly1 and Munc18c suggested a united, positive impact upon SNARE‐based fusion of ER‐to‐Golgi‐ and Golgi‐to‐plasma membrane‐addressed exocytic vesicles and increased the secretory capacity of different therapeutic proteins in Chinese hamster ovary cells up to 40 pg/cell/day. Sly1‐ and Munc18c‐based vesicle traffic engineering cooperated with Xbp‐1‐mediated ER/Golgi organelle engineering. Our study supports a model for united function of SM proteins in stimulating vesicle trafficking machinery and provides a generic secretion engineering strategy to improve biopharmaceutical manufacturing of important protein therapeutics. Biotechnol. Bioeng. 2009;102: 1170‐1181. © 2008 Wiley Periodicals, Inc.