Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation.
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.
Jung
Yuchul
Y
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.
Rhee
Yong
Y
Auh
Chung-Kyoon
CK
Shim
Hyekyung
H
Choi
Jung-Jin
JJ
Kwon
Suk-Tae
ST
Yang
Joo-Sung
JS
Kim
Donggiun
D
Kwon
Myung-Hee
MH
Kim
Yong-Sung
YS
Lee
Sukchan
S
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
D007135:Immunoglobulin Variable Region; D018749:RNA, Plant
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