[Biosynthesis of mini-antibodies to human ferritin in plant and bacterial producers].
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.
Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow Region, 142290 Russia. centaurea@mail.ru
E G
EG
Semeniuk
O A
OA
Stremovskiĭ
I V
IV
Orlova
T G
TG
Baladin
A M
AM
Nosov
Ia I
IaI
Bur'ianov
S M
SM
Deev
R V
RV
Petrov
D001419:Bacteria / Q000276:immunology*; D001426:Bacterial Proteins; D001483:Base Sequence; D002846:Chromatography, Affinity; D017931:DNA Primers; D005293:Ferritins / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis*; D010944:Plants / Q000276:immunology*; D011506:Proteins / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry
D001426:Bacterial Proteins; D017931:DNA Primers; D007128:Immunoglobulin Fragments; D011506:Proteins; D011993:Recombinant Fusion Proteins; C057438:barstar protein, Bacillus amyloliquefaciens; D005293:Ferritins; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease