Expression of anti-breast cancer monoclonal antibody in transgenic plant
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
BACKGROUND: Plant expression system for mass production of recombinant proteins has several advantages over other existing expression systems with economical and safety issues. Breast cancer is a cancer originating from breast tissue and comprises almost 25% of all cancers in women world widely. Lewis-Y antigen is difucosylated oligosaccharide and is carried by glycoconjugates at cancer cell surface. In this study, the anti-breast cancer mAb BR55, which recognizes the epitope Lewis-Y, was expressed in the plant expression system. METHODS AND RESULTS:We have developed plant system for production of mAb BR55 with or without KDEL (the ER retention signal). This ER retention signal was attached to C-terminus of protein to help retain the recombinant glycoprotein carrying oligomannose glycans and enhance glycoprotein accumulation. PCR analysis was performed and confirmed the presence of recombinant genes. Western blot validated that the recombinant proteins mAb BR55 with or without KDEL were expressed in transgenic plants, moreover, the expression level of the mAb BR55 with KDEL was higher compared to the mAb BR55 without KDEL. CONCLUSION:These results indicate that KDEL fusion is a good way to produce proteins and plant can be an ideal expression system to obtain proteins and enhance accumulation of proteins.
Abstract
BACKGROUND: Plant expression system for mass production of recombinant proteins has several advantages over other existing expression systems with economical and safety issues. Breast cancer is a cancer originating from breast tissue and comprises almost 25% of all cancers in women world widely. Lewis-Y antigen is difucosylated oligosaccharide and is carried by glycoconjugates at cancer cell surface. In this study, the anti-breast cancer mAb BR55, which recognizes the epitope Lewis-Y, was expressed in the plant expression system. METHODS AND RESULTS:We have developed plant system for production of mAb BR55 with or without KDEL (the ER retention signal). This ER retention signal was attached to C-terminus of protein to help retain the recombinant glycoprotein carrying oligomannose glycans and enhance glycoprotein accumulation. PCR analysis was performed and confirmed the presence of recombinant genes. Western blot validated that the recombinant proteins mAb BR55 with or without KDEL were expressed in transgenic plants, moreover, the expression level of the mAb BR55 with KDEL was higher compared to the mAb BR55 without KDEL. CONCLUSION:These results indicate that KDEL fusion is a good way to produce proteins and plant can be an ideal expression system to obtain proteins and enhance accumulation of proteins.
참고문헌 (19)
- Brodzik, R, "Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells", Pro Natl Acad Sci USA, 103, pp.8804~8809 (2006)
- Wandelt, C.I., "Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants", The Plant J, 2, pp.181~192 (1992)
- Dettke, M., "Activationdependent expression of the blood group-related lewis Y antigen on peripheral blood granulocytes", J. Leukocyte Biol, 68, pp.511~514 (2000)
- Fernandes, B., "Beta 1-6 branched oligosaccharides as a marker of tumor progression in human breast and colon neoplasia", Cancer Res, 51, pp.718~723 (1991)
- Gomord, V, "qThe C-terminal HDEL sequence is sufficient for retention of secretory proteins in the endoplasmic reticulum (ER) but promotes vacuolar targeting of proteins that escape the ER", Plant J, 11, pp.313~325 (1997)
- Góra-Sochacka, A., "Plant-based production of recombinant cytokines", Postepy Biochem, 55, pp.85~94 (2009)
- Ko, K., "Plant biopharming of monoclonal antibodies", Virus Res, 111, pp.93~100 (2005)
- Ko, K., "Effect of untranslated leader sequence of AMV RNA 4 and signal peptide of pathogenesisrelated protein 1b on attacin gene expression, and resistance to fire blight in transgenic apple", Biotechnol. Lett, 22, pp.373~381 (2000)
- Ko, K., "Inhibition of tumor growth by plant-derived mAb", Proc Natl Acad Sci USA, 102, pp.7026~7030 (2005)
- Ko, K, "Function and glycosylation of plant-derived antiviral monoclonal antibody", Pro Natl Acad Sci USA, 100, pp.8013~8018 (2003)
- Koprowski, H., "Vaccines and sera through plant biotechnology", Vaccine, 23, pp.1757~1763 (2005)
- Nemoto-Sasaki, Y, "Correlation between the sialylation of cell surface Thomsen-Friedenreich antigen and the metastatic potential of colon carcinoma cells in a mouse model", Glycoconj J, 18, pp.895~906 (2001)
- Nichols, E.J, "Carbohydrate determinants associated with carcinoembryonic antigen (CEA)", J. Immunol, 135, pp.1911~1913 (1985)
- Pagny, S, "Structural requirements for Arabidopsis β 1,2-xylosyltransferase activity and targeting to the Golgi", The Plant Journal, 33, pp.189~203 (2003)
- Schillberg, S., "Molecular farming of recombinant antibodies in plants", Cell Mol Life Sci, 60, pp.433~445 (2003)
- Schouten, A., "The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco", Plant Mol Biol, 30, pp.781~793 (1996)
- Steplewski, Z., "Tumor cell lysis and tumor growth inhibition by the isotype variants of MAb BR55-2 directed against Y oligosaccharide", In Vivo, 5, pp.79~83 (1991)
- Stoger, E, "Plantibodies: applications, advantages and bottlenecks", Curr Opin Biotechnol, 13, pp.161~166 (2002)
- Wandelt, C.I., "Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants", The Plant J, 2, pp.181~192 (1992)