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Multiple Monoclonal Antibodies Produced in a Single Transgenic Plant

형질전환 식물체에서의 복합 단일 항체 단백질 생산

  • Published : 2009.01.30

Abstract

Production of highly valuable immunotherapeutic proteins such as monoclonal antibodies and vaccines using plant biotechnology and genetic engineering has been studied as a popular research field. Plant expression system for mass production of such useful recombinant therapeutic proteins has several advantages over other existing expression systems with economical and safety issues. Immunotherapy of multiple monoclonal antibodies, which can recognize multiple targeting including specific proteins and their glycans highly expressed on the surface of cancer cells, can be an efficient treatment compared to a single targeting immunotherapy using a single antibody. In this study, we have established plant production system to express two different targeting monoclonal antibodies in a single transgenic plant through crossing fertilization between two different transgenic plants expressing anti-colorectal cancer mAbCO17-1A and anti-breast cancer mAbBR55, respectively. The F1 seedlings were obtained cross fertilization between the two transgenic parental plants. The presence, transcription, and protein expression of heavy chain (HC) and light chain (LC) genes of both mAbs in the seedlings were investigated by PCR, RT-PCR, and immunoblot analyses, respectively. Among all the seedlings, some seedlings did not carry or transcribe the HC and LC genes of both mAbs. Thus, the seedlings with presence and transcription of HC and LC genes of both mAbs were selected, and the selected seedlings were confirmed to have relatively stronger density of HC and LC protein bands compared to the transgenic plant expressing only each mAb. These results indicate that the F1 seedling plant with carrying both mAb genes was established. Taken together, plant crossing fertilization can be applied to generate an efficient production system expressing multiple monoclonal antibodies for immunotherapy in a single plant.

Keywords

Crossing fertilization;mAb BR55;mAb CO17-1A

References

  1. Adams, G. P. and L. M. Weiner. 2005. Monoclonal antibody therapy of cancer. Nat. Biotechnol. 23, 1147-1157 https://doi.org/10.1038/nbt1137
  2. Agarwal, S., R. Singh, I. Sanyal and D. V. Amla. 2008. Expression of modified gene encoding functional human alpha-1-antitrypsin protein in transgenic tomato plants. Transgenic Res. [Epub ahead of print]
  3. Bauly, J. M., I. M. Sealy, H. Macdonald, J. Brearley, S. Droge, S. Hillmer, D. G. Robinson, M. A. Venis, M. R. Blatt, C. M. Lazarus and R. M. Napier. 2000. Overexpression of auxin-binding protein enhances the sensitivity of guard cells to auxin. Plant Physiol. 124, 1229-1238 https://doi.org/10.1104/pp.124.3.1229
  4. Brodzik, R., M. Glogowska, K. Bandurska, M. Okulicz, D. Deka, K. Ko. J. van der Linden, J. H. Leusen, N. Pogrebnyak, M. Golovkin, Z. Steplewski and H. Koprowski. 2006. Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells. Proc. Natl. Acad. Sci. USA 103, 8804-8809 https://doi.org/10.1073/pnas.0603043103
  5. Flieger, D., A. S. Hoff, T. Sauerbruch and I. G. Schmidt- Wolf. 2001. Influence of cytokines, monoclonal antibodies and chemotherapeutic drugs on epithelial cell adhesion molecule (EpCAM) and LewisY antigen expression. Clin. Exp. Immunol. 123, 9-14 https://doi.org/10.1046/j.1365-2249.2001.01435.x
  6. Helenius, A. and M. Aebi. 2001. Intracellular functions of N-linked glycans. Science 291, 2364-2369 https://doi.org/10.1126/science.291.5512.2364
  7. Ko, K., Z. Steplewski, M. Glogowska and H. Koprowski. 2005. Inhibition of tumor growth by plant-derived mAb. Proc. Natl. Acad. Sci. USA 102, 7026-7030 https://doi.org/10.1073/pnas.0502533102
  8. Ko, K., Y. Tekoah, P. M. Rudd, D. J. Harvey, R. A. Dwek, S. Spitsin, C. A. Hanlon, C. Rupprecht, B. Dietzschold, M. Golovkin and H. Koprowski. 2003. Function and glycosylation of plant-derived antiviral monoclonal antibody. Proc. Natl. Acad. Sci. USA 100, 8013-8018 https://doi.org/10.1073/pnas.0832472100
  9. Luo, P., M. Agadjanyan, J. Qiu, M. A. Westerink, Z. Steplewski and T. Kieber-Emmons. 1998. Antigenic and immunological mimicry of peptide mimotopes of Lewis carbohydrate antigens. Mol. Immunol. 35, 865-879 https://doi.org/10.1016/S0161-5890(98)00067-4
  10. Ma, J. K., P. M. Drake and P. Christou. 2003. The production of recombinant pharmaceutical proteins in plants. Nat. Rev. Genet. 4, 794-805 https://doi.org/10.1038/nrg1177
  11. Madjd, Z., T. Parsons, N. F. Watson, I. Spendlove, I. Ellis and L. G. Durrant. 2005. High expression of Lewis y/b antigens is associated with decreased survival in lymph node negative breast carcinomas. Breast Cancer Res. 7, R780-87 https://doi.org/10.1186/bcr1305
  12. Muldoon, L. L. and E. A. Neuwelt. 2003. BR96-DOX immunoconjugate targeting of chemotherapy in brain tumor models. J. Neurooncol. 65, 49-62 https://doi.org/10.1023/A:1026234130830
  13. Nemoto-Sasaki, Y., M. Mitsuki, M. Morimoto-Tomita, A. Maeda, M. Tsuiji and T. Irimura. 2001. 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, 895-906 https://doi.org/10.1023/A:1022252509765
  14. Pai-Scherf, L. H., J. A. Carrasquillo, C. Paik, O. Gansow, M. Whatley, D. Pearson, K. Webber, M. Hamilton, C. Allegra, M. Brechbiel, M. C. Willingham and I. Pastan. 2000. Imaging and phase I study of 111In- and 90Y-labeled anti-LewisY monoclonal antibody B3. Clin. Cancer Res. 6, 1720-1730
  15. Plunkett, T. A. and D. W. Miles. 2002. New biological therapies for breast cancer. Int. J. Clin. Pract. 56, 261-266
  16. Richter, L. J., Y. Thanavala, C. J. Arntzen and H. S. Mason. 2000. Production of hepatitis B surface antigen in transgenic plants for oral immunization. Nat. Biotechnol. 18, 1167-1171 https://doi.org/10.1038/81153
  17. Scherf, U., D. T. Ross, M. Waltham, L. H. Smith, J. K. Lee, L. Tanabe, K. W. Kohn, W. C. Reinhold, T. G. Myers, D. T. Andrews, D. A. Scudiero, M. B. Eisen, E. A. Sausville, Y. Pommier, D. Botstein, P. O. Brown and J. N. Weinstein. 2000. A gene expression database for the molecular pharmacology of cancer. Nat. Genet. 24, 236-244 https://doi.org/10.1038/73439
  18. Tacket, C. O., H. S. Mason, G. Losonsky, J. D. Clements, M. M. Levine and C. J. Arntzen. 1998. Immunogenicity in humans of a recombinant bacterial antigen delivered in a transgenic potato. Nat. Med. 4, 607-609 https://doi.org/10.1038/nm0598-607
  19. Youm, J. W., J. H. Jeon, H. Kim, Y. H. Kim, K. Ko, H. Joung and H. Kim. 2008. Transgenic tomatoes expressing human beta-amyloid for use as a vaccine against Alzheimer's disease. Biotechnol. Lett. [Epuib ahead of print]
  20. Zaloudik, J., W. Li, L. Jacob, M. P. Kieny, R. Somasundaram, B. Acres, H. Song, T. Zhang, J. Li and D. Herlyn. 2002. Inhibition of tumor growth by recombinant vaccinia virus expressing GA733/CO17-1A/EpCAM/KSA/KS1-4 antigen in mice. Cancer Gene Ther. 9, 382-389 https://doi.org/10.1038/sj.cgt.7700452