title: Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation.
abstract: We developed an asexual reproductive plant, Kalanchoe pinnata, as a new bioreactor for plant-based molecular farming using a newly developed transformation method. Leaf crenate margins were pin-pricked to infect the plant with the Agrobacterium strain LBA4404 and vacuum infiltration was also applied to introduce the target gene into the plants. Subsequently, the young mother leaf produced new clones at the leaf crenate margins without the need for time- and labor-consuming tissue culture procedures. The average transformation rates were approximately 77 and 84% for pin-prickling and vacuum-infiltration methods, respectively. To functionally characterize an introduced target protein, a nucleic acid hydrolyzing recombinant 3D8 scFv was selected and the plant based 3D8 scFv proteins were purified and analyzed. Based on abzyme analysis, the purified protein expressed with this system had catalytic activity and exhibited all of properties of the protein produced in an E. coli system. This result suggested that vegetatively reproductive K. pinnata can be a novel and potent bioreactor for bio-pharmaceutical proteins.
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.
authors.forename: Yuchul
authors.initials: Y
authors.lastname: Jung
authors.forename: Yong
authors.initials: Y
authors.lastname: Rhee
authors.forename: Chung-Kyoon
authors.initials: CK
authors.lastname: Auh
authors.forename: Hyekyung
authors.initials: H
authors.lastname: Shim
authors.forename: Jung-Jin
authors.initials: JJ
authors.lastname: Choi
authors.forename: Suk-Tae
authors.initials: ST
authors.lastname: Kwon
authors.forename: Joo-Sung
authors.initials: JS
authors.lastname: Yang
authors.forename: Donggiun
authors.initials: D
authors.lastname: Kim
authors.forename: Myung-Hee
authors.initials: MH
authors.lastname: Kwon
authors.forename: Yong-Sung
authors.initials: YS
authors.lastname: Kim
authors.forename: Sukchan
authors.initials: S
authors.lastname: Lee
mesh_terms: D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000302:isolation & purification; D031962:Kalanchoe / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics; D012231:Rhizobium / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D007135:Immunoglobulin Variable Region; D018749:RNA, Plant
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