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Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation

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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.

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Acknowledgments

This study was supported in part by a grant from Rural Development Administration in Korea (BioGreen21, 20080701-034-001-008-02-00) and by a grant from National Veterinary Research and Quarantine Service, Ministry of Food, Agriculture, Forestry, and Fisheries in 2008 (Project Code No., Z-AD14-2008-08-03), and Korean Research Foundation grant (KRF 2003-005-D00010).

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Authors and Affiliations

  1. Department of Genetic Engineering, Sungkyunkwan University, Suwon, 440-746, Korea

    Yuchul Jung, Yong Rhee, HyeKyung Shim, Jung-Jin Choi, Suk-Tae Kwon, Joo-Sung Yang, Donggiun Kim & Sukchan Lee

  2. Division of Life Sciences, Mokpo National University, Muan, 534-729, Korea

    Chung-Kyoon Auh

  3. Department of Microbiology, Ajou University School of Medicine, Suwon, 443-721, Korea

    Myung-Hee Kwon

  4. Department of Molecular Science and Technology, Ajou University, Suwon, 443-749, Korea

    Yong-Sung Kim

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  1. Yuchul Jung
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  2. Yong Rhee
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  3. Chung-Kyoon Auh
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  4. HyeKyung Shim
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  5. Jung-Jin Choi
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  7. Joo-Sung Yang
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  8. Donggiun Kim
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  9. Myung-Hee Kwon
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  10. Yong-Sung Kim
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  11. Sukchan Lee
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Corresponding author

Correspondence to Sukchan Lee.

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Communicated by J. R. Liu.

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Jung, Y., Rhee, Y., Auh, CK. et al. Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation. Plant Cell Rep 28, 1593–1602 (2009). https://doi.org/10.1007/s00299-009-0758-3

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  • DOI: https://doi.org/10.1007/s00299-009-0758-3

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