Production of functional human nerve growth factor from the saliva of transgenic mice by using salivary glands as bioreactors

利用唾液腺作为生物反应器从转基因小鼠唾液中生产功能性人神经生长因子

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作者:Fang Zeng, Zicong Li, Qingchun Zhu, Rui Dong, Chengcheng Zhao, Guoling Li, Guo Li, Wenchao Gao, Gelong Jiang, Enqin Zheng, Gengyuan Cai, Stefan Moisyadi, Johann Urschitz, Huaqiang Yang, Dewu Liu, Zhenfang Wu

Abstract

The salivary glands of animals have great potential to act as powerful bioreactors to produce human therapeutic proteins. Human nerve growth factor (hNGF) is an important pharmaceutical protein that is clinically effective in the treatment of many human neuronal and non-neuronal diseases. In this study, we generated 18 transgenic (TG) founder mice each carrying a salivary gland specific promoter-driven hNGF transgene. A TG mouse line secreting high levels of hNGF protein in its saliva (1.36 μg/mL) was selected. hNGF protein was successfully purified from the saliva of these TG mice and its identity was verified. The purified hNGF was highly functional as it displayed the ability to induce neuronal differentiation of PC12 cells. Furthermore, it strongly promoted proliferation of TF1 cells, above the levels observed with mouse NGF. Additionally, saliva collected from TG mice and containing unpurified hNGF was able to significantly enhance the growth of TF1 cells. This study not only provides a new and efficient approach for the synthesis of therapeutic hNGF but also supports the concept that salivary gland from TG animals is an efficient system for production of valuable foreign proteins.

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