Transient plant production of Salmonella Typhimurium diagnostic antibodies

Kopertekh, Lilya; Meyer, Torsten; Freyer, Cornelia; Hust, Michael

The biological activity of plant-generated antibodies is usually compared to that of reference antibodies produced in bacterial or animal cell expression systems. Therefore, the scFv-TM43-E10 and scFv-Fc-TM43-E10 antibody derivatives have been also expressed in bacterial and mammalian cell cultures and included into the 7 . ELISA test of scFv-TM43-E10 and scFv-Fc-TM43-E10 antibody fragments. ELISA plates were coated with S. Typhimurium suspension (lane 1), protein extracts from S. Typhimurium infected (lanes 2-5) and non-infected (lane 6) salad leaf tissue. 400 ng/well for scFv-TM43-E10 and 150 ng/well for scFv-Fc-TM43-E10 were added for detection of OmpD antigen. Values represent the means with standard deviation from two independent tests. functional test. The antigen binding of plant-derived scFv-TM43-E10 antibodies was similar to scFv-TM43-E10 recombinant proteins isolated from E. coli. Furthermore functional ELISA test also showed a similar functional activity for plant and mammalian cell culture-made scFv-Fc-TM43-E10 antibody fragments (Fig. 5A ). Our data are in agreement with findings described in several articles with respect to the functional equivalence of the recombinant proteins produced in different expression systems. For example plant-derived 6D8 and hE16 mAbs demonstrated similar to mammalian cell-derived counterpart antigen specificity [58] . Yuan et al. [59] reported that the biological activity of antizearalenone scFv "plantibody" was comparable to that of a bacterially produced scFv antibody. A number of research groups investigated the binding properties of the plant-derived anti-HIV VRC01, 2G12 and b12 antibodies in comparison to reference counterparts [60] [61] [62] . These studies also revealed functional equivalence between plant-and mammalian cells-made recombinant proteins.