Expression of single-chain antibody-barstar fusion in plants |
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| Authors: | Semenyuk Ekaterina G Stremovskiy Oleg A Edelweiss Evelina F Shirshikova Olga V Balandin Taras G Buryanov Yaroslav I Deyev Sergey M |
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| Institution: | Branch of Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Nauki Avenue, Pushchino, Moscow region 142290, Russia. |
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| Abstract: | We successfully cloned and expressed a single-chain antibody (425scFv), that is directed to human epidermal growth factor receptor HER1 (EGFR) in transgenic tobacco plants as a fusion with bacterial barstar gene (425scFv-barstar). Plant-produced recombinant 425scFv-barstar was recovered using barstar-barnase system. Based on barstar-barnase affinity, during purification of the plant-produced 425scFv-barstar, we generated bispecific scFv-antibody heterodimers from individual single-chain fragments initially produced in different host systems with binding activity to both HER1 and HER2/neu tumor antigens. We demonstrated by flow cytometry and indirect immunofluorescent microscopy that both the components of heterodimer retain its specific cell-binding activity. |
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| Keywords: | Transgenic plants Single-chain Fv antibody Barstar Barnase HER1 (EGFR) |
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