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A fusion‐protein approach enabling mammalian cell production of tumor targeting protein domains for therapeutic development |
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| Authors: | Jia Hu Xiang Chen Xuhua Zhang Xiaopeng Yuan Mingjuan Yang Hui Dai Wei Yang Qinghua Zhou Weihong Wen Qirui Wang Weijun Qin Aizhi Zhao |
| Affiliation: | 1. Lung Cancer Research Center, West China Hospital, Sichuan University, Chengdu, China;2. Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania;3. School of Life Sciences, Zhengzhou University, Zhengzhou, China;4. Zhujiang Hospital, SouthernMedical University, Guangzhou, China;5. Xinjiang Karamay Central Hospital, Karamay City, Xinjiang, China;6. State Key Laboratory of Cancer Biology, Department of Immunology, Xijing Hospital, Fourth Military Medical University, China, Xi'an;7. College of Traditional Chinese Medicine, SouthernMedical University, Guang Dong, China;8. Department of Urology, Xijing Hospital, Fourth Military Medical University, China, Xi'an |
| Abstract: | A single chain Fv fragment (scFv) is a fusion of the variable regions of heavy (VH) and light (VL) chains of immunoglobulins. They are important elements of chimeric antigen receptors for cancer therapy. We sought to produce a panel of 16 extracellular protein domains of tumor markers for use in scFv yeast library screenings. A series of vectors comprising various combinations of expression elements was made, but expression was unpredictable and more than half of the protein domains could not be produced using any of the constructs. Here we describe a novel fusion expression system based on mouse TEM7 (tumor endothelial marker 7), which could facilitate protein expression. With this approach we could produce all but one of the tumor marker domains that could not otherwise be expressed. In addition, we demonstrated that the tumor associated antigen hFZD10 produced as a fusion protein with mTEM7 could be used to enrich scFv antibodies from a yeast display library. Collectively our study demonstrates the potential of specific fusion proteins based on mTEM7 in enabling mammalian cell production of tumor targeting protein domains for therapeutic development. |
| Keywords: | recombinant protein expression scFv mammalian cells cancer yeast library screening antibodies |