Foot-and-mouth disease virus (FMDV) is the causative agent of an economically important disease that affects cattle and other clovenhoofed animals. VP1 is the most frequently studied protein owing to its significant roles in virus attachment, protective immunity, and serotype specificity. Here, we report high cell density cultivations of Escherichia coli for preparative scale production of FMDV VP1 in 5 L bioreactor. The epitope gene was expressed in cytoplasm of E. coli as GST fused format and its soluble expression (~ 90 %) was first confirmed by flask cultivation. After simple purification, antigenic activity of VP1 epitope was validated by western blot analysis and ELISA with commercially available monoclonal antibody. For large scale production pH-stat fedbatch cultivation was performed in 5 L jar bioreactor and various conditions were examined for higher productivities. Among examined conditions, when cells were induced at higher cell density (OD600=100) and complex feeding solutions were supplemented, significantly higher production yields could be achieved. The produced VP1 epitope can be used for development of protein therapeutics as well as diagnostic system.
키워드
foot-and-mouth disease virusVP1 epitopehigh-cell density cultivation
저자
Joon-Goo JUNG [ Dept. of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon 305-701. ]
Ki Jun JEONG [ Dept. of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon 305-701. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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