Expression of single-chain antibody-barstar fusion in plants

Expression of single-chain antibody-barstar fusion in plants

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 barsta...

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Veröffentlicht in:Biochimie 2007, Vol.89 (1), p.31-38
Hauptverfasser: Semenyuk, Ekaterina G., Stremovskiy, Oleg A., Edelweiss, Evelina F., Shirshikova, Olga V., Balandin, Taras G., Buryanov, Yaroslav I., Deyev, Sergey M.
Format: Artikel
Sprache:eng
Schlagworte:
Bacterial Proteins - genetics
Barnase
Barstar
Blotting, Western
Cloning, Molecular
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
ErbB Receptors - immunology
Flow Cytometry
HER1 (EGFR)
Humans
Immunoglobulin Fragments - biosynthesis
Immunoglobulin Fragments - immunology
Microscopy, Fluorescence
Nicotiana - genetics
Plants, Genetically Modified
Protein Engineering - methods
Receptor, ErbB-2
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - immunology
Single-chain Fv antibody
Transgenic plants
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Zusammenfassung: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.
ISSN:0300-9084
1638-6183
DOI:10.1016/j.biochi.2006.07.012