Anti-solasodine glycoside single-chain Fv antibody stimulates biosynthesis of solasodine glycoside in plants

• Published in: Plant cell reports Vol. 22; no. 5; pp. 344 - 349
• Main Authors: PUTALUN, W, TAURA, F, QING, W, MATSUSHITA, H, TANAKA, H, SHOYAMA, Y
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.12.2003; Springer Nature B.V
• Subjects: Amino Acid Sequence; Antibodies - genetics; Antibodies - metabolism; Antineoplastic drugs. Cytokines; Bacteria; Base Sequence; Biological and medical sciences; Biotechnology; Cell Line; E coli; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant; Glycosides - biosynthesis; Glycosides - chemistry; Glycosides - immunology; Health. Pharmaceutical industry; Hybridomas; Immunoglobulin Fragments - immunology; Industrial applications and implications. Economical aspects; Molecular Sequence Data; Plant Proteins - genetics; Plant Proteins - metabolism; Plant Roots - genetics; Plant Roots - metabolism; Plants, Genetically Modified; Production of active biomolecules; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Solanaceous Alkaloids - biosynthesis; Solanaceous Alkaloids - chemistry; Solanaceous Alkaloids - genetics; Solanaceous Alkaloids - immunology; Solanaceous Alkaloids - metabolism; Solanum - genetics; Solanum - metabolism; Solanum khasianum; Antineoplastic agent; Drug; Hairy root; Solasodine glycosides; Bioactive material; Biosynthesis; Metabolic pathway; Medicinal plant; Solanum khasianum; Alkaloid; Plant breeding; Transgenic hairy root; scFv; Dicotyledones; Angiospermae; Production; Spermatophyta; Chemical concentration; Glycoside; Transgenic plant; Yield; Solanaceae; Single chain antibody
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-003-0689-3

We constructed a recombinant antibody fragment--single chain fragment-variable (scFv) antibody--derived from hybridoma cell lines to control the concentration of solasodine glycosides in hairy root cultures of Solanum khasianum transformed by the anti-solamargine (As)-scFv gene. The properties of the As-scFv protein expressed in Escherichia coli were almost identical to those of the parent monoclonal antibody (MAb). Up to 220 ng recombinant As-scFv was expressed per milligram of soluble protein in transgenic hairy root cultures of S. khasianum. The concentration of solasodine glycosides was 2.3-fold higher in the transgenic than in the wild-type hairy root, as reflected by the soluble As-scFv level and antigen binding activities. These results suggested that the scFv antibody expressed in transgenic hairy roots controlled the antigen level, thus representing a novel plant breeding methodology that can produce secondary metabolites.