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 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.
Semenyuk
Ekaterina G
EG
Branch of Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Nauki Avenue, Pushchino, Moscow region 142290, Russia.
Stremovskiy
Oleg A
OA
Edelweiss
Evelina F
EF
Shirshikova
Olga V
OV
Balandin
Taras G
TG
Buryanov
Yaroslav I
YI
Deyev
Sergey M
SM
D001426:Bacterial Proteins / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D066246:ErbB Receptors / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000276:immunology; D008856:Microscopy, Fluorescence; D030821:Plants, Genetically Modified; D015202:Protein Engineering / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D014026:Nicotiana / Q000235:genetics*
D001426:Bacterial Proteins; D066246:ErbB Receptors; D007128:Immunoglobulin Fragments; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; C057438:barstar protein, Bacillus amyloliquefaciens; C512478:EGFR protein, human