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Biosynthesis of the scFv Antibody to Human Ferritin in Plant and Bacterial Producers |
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| Authors: | Semenyuk E. G. Stremovskii O. A. Orlova I. V. Balandin T. G. Nosov A. M. Buryanov Ya. I. Deyev S. M. Petrov R. V. |
| Affiliation: | (1) Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia;(2) Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia;(3) Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia;(4) Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, 127276, Russia |
| Abstract: | A recombinant scFv antibody against human spleen ferritin was expressed as a barstar-fused protein in Escherichia coli and in Nicotiana tabacum plants and suspension cell cultures. As demonstrated by immunoblotting with antibarstar antibodies, direction of the recombinant protein to the endomembrane system of plant cells ensured its stability and solubility. Production of the recombinant protein did not differ between parental transgenic plants and their first-generation progeny. Fusion with barstar allowed not only immunochemical detection of the recombinant scFv antibody, but also their purification from the plant material by affinity chromatography with barnase-His6 immobilized on a metal-affinity carrier. |
| Keywords: | transgenic plants variable antibody domains barstar barnase |
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