Bacterial and plant-produced scFv proteins have similar antigen-binding properties.
A gene encoding a single-chain variable (scFv) antibody fragment was expressed as a cytoplasmic and endoplasmic reticulum-targeted protein in transgenic tobacco plants. In both cases, the scFv accumulated up to 0.01% of total soluble protein (TSP). The same scFv fragment was also produced in the periplasm of Escherichia coli. Measurement of the affinity by ELISA indicates that the affinity of the bacterially made scFv is about 80-fold lower than that of the parental Fab fragment. The results suggest that the affinity of the plant-produced scFv fragments is reduced to a similar extent, implying that all the plant-produced scFv fragments are antigen binding.
Bruyns
A M
AM
Department of Genetics, Flanders Interuniversity Institute for Biotechnology, Universiteit Gent, Belguim.
De Jaeger
G
G
De Neve
M
M
De Wilde
C
C
Van Montagu
M
M
Depicker
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D003593:Cytoplasm / Q000276:immunology; D004721:Endoplasmic Reticulum / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology; D005822:Genetic Vectors; D015151:Immunoblotting; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis / Q000276:immunology; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011817:Rabbits; D014026:Nicotiana / Q000235:genetics / Q000276:immunology*
D001426:Bacterial Proteins; D007128:Immunoglobulin Fragments; D010940:Plant Proteins; C055966:immunoglobulin Fv