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Production of monoclonal antibodies against antitumor cyclohexapeptide RA-VII from Rubia cordifolia and their characterization

Journal of Natural Medicines volume 65, pages 588–593 (2011)Cite this article

Abstract

An immunoassay system was established for the estimation of the quantity of an antitumor cyclic hexapeptide RA-VII (1) from Rubia cordifolia L. and R. akane Nakai (Rubiaceae). First, 1 was converted into its hapten, which was then conjugated with a carrier protein to be used as an effective antigen to obtain its monoclonal antibody (MAb). In the resulting conjugate, the molecular ratio between 1 and the carrier protein as assayed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was about 5:1. Then, the splenocytes from the mouse immunized with the conjugate were fused with mouse myeloma cells to produce hybridoma, secreting MAb against 1. Two clones were isolated, one producing MAb IgG1 and the other IgM, both having a κ light chain. The sensitivity and cross-reactivity of the thus obtained MAb were also assayed.

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Fig. 1
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References

  1. Itokawa H, Takeya K, Mihara K, Mori N, Hamanaka T, Sonobe T, Iitaka Y (1983) Studies on the antitumor cyclic hexapeptides obtained from Rubiae radix. Chem Pharm Bull 31:1424–1427

    PubMed  CAS  Google Scholar 

  2. Itokawa H, Takeya K, Mori N, Hamanaka T, Sonobe T, Mihara K (1984) Isolation and antitumor activity of cyclic hexapeptides isolated from Rubiae radix. Chem Pharm Bull 32:284–290

    PubMed  CAS  Google Scholar 

  3. Itokawa H, Takeya K, Hitotsuyanagi Y, Morita H (1997) Macrocyclic peptide alkaloids from plants. In: Cordell GA (ed) The alkaloids, vol 49. Academic Press, New York, pp 301–387

    Google Scholar 

  4. Jolad SD, Hoffmann JJ, Torrance SJ, Wiedhopf RM, Cole JR, Arora SK, Bates RB, Gargiulo RL, Kriek GR (1977) Bouvardin and deoxybouvardin, antitumor cyclic hexapeptides from Bouvardia ternifolia (Rubiaceae). J Am Chem Soc 99:8040–8044. doi:10.1021/ja00466a043

    PubMed  Article  CAS  Google Scholar 

  5. Inoue K, Mukaiyama T, Kobayashi T, Ogawa M (1986) Activity of RA-700, a cyclic hexapeptide from Rubiae Radix, in the human tumor clonogenic assay. Invest New Drugs 4:231–236. doi:10.1007/BF00179588

    PubMed  Article  CAS  Google Scholar 

  6. Majima H, Tukagoshi S, Furue H, Suminaga M, Sakamoto K, Nukaria N, Uematsu K, Furuta T, Kurihara M, Yoshida A, Isomura S, Takemoto T, Hirashima M, Izumi T, Nakao I, Ohashi Y, Ito K, Asai R (1993) Jpn J Cancer Chemother 20:67–78

    CAS  Google Scholar 

  7. Zalacaín M, Zaera E, Vázquez D, Jiménez A (1982) The mode of action of the antitumor drug bouvardin, an inhibitor of protein synthesis in eukaryotic cells. FEBS Lett 148:95–97. doi:10.1016/0014-5793(82)81250-7

    PubMed  Article  Google Scholar 

  8. Sirdeshpande BV, Toogood PL (1995) Inhibition of protein synthesis by RA-VII. Bioorg Chem 23:460–470. doi:10.1006/bioo.1995.1032

    Article  CAS  Google Scholar 

  9. Fujiwara H, Saito S, Hitotsuyanagi Y, Takeya K, Ohizumi Y (2004) RA-VII, a cyclic depsipeptide, changes the conformational structure of actin to cause G2 arrest by the inhibition of cytokinesis. Cancer Lett 209:223–229. doi:10.1016/j.canlet.2003.12.022

    PubMed  Article  CAS  Google Scholar 

  10. Sakata R, Shoyama Y, Murakami H (1994) Production of monoclonal antibodies and enzyme immunoassay for typical adenylate cyclase activator, forskolin. Cytotechnology 16:101–108. doi:10.1007/BF00754612

    PubMed  Article  CAS  Google Scholar 

  11. Yanagihara H, Sakata R, Shoyama Y, Murakami H (1996) Rapid analysis of small samples containing forskolin using monoclonal antibodies. Planta Med 62:169–172

    PubMed  Article  CAS  Google Scholar 

  12. Shoyama Y, Fukada T, Murakami H (1996) Production of monoclonal antibodies and ELISA for thebaine and codeine. Cytotechnology 19:55–61. doi:10.1007/BF00749755

    Article  Google Scholar 

  13. Tanaka H, Goto Y, Shoyama Y (1996) Monoclonal antibody based enzyme immunoassay for marihuana (cannabinoid) compounds. J Immunoassay 17:321–342

    PubMed  Article  CAS  Google Scholar 

  14. Ishiyama M, Shoyama Y, Murakami H, Shinohara H (1996) Production of monoclonal antibodies and development of an ELISA for solamargine. Cytotechnology 18:153–158. doi:10.1007/BF00767762

    Article  Google Scholar 

  15. Itokawa H, Takeya K, Mori N, Sonobe T, Mihashi S, Hamanaka T (1986) Studies on antitumor cyclic hexapeptides RA obtained from Rubiae Radix, Rubiaceae. VI. Minor antitumor constituents. Chem Pharm Bull 34:3762–3768

    PubMed  CAS  Google Scholar 

  16. Itokawa H, Morita H, Takeya K, Tomioka N, Itai A, Iitaka Y (1991) New antitumor bicyclic hexapeptides, RA-VI and -VIII from Rubia cordifolia; conformation–activity relationship II. Tetrahedron 47:7007–7020. doi:10.1016/S0040-4020(01)96155-1

    Article  CAS  Google Scholar 

  17. Morinaga O, Tanaka H, Shoyama Y (2000) Production of monoclonal antibody against a major purgative component, sennoside A, its characterization and ELISA. Analyst 125:1109–1113. doi:10.1039/B000988L

    Article  CAS  Google Scholar 

  18. Weiler EW, Zenk MH (1976) Use of immunoassay in plant science. Part 1. Radioimmunoassay for the determination of digoxin and related compounds in Digitalis lanata. Phytochemistry 15:1537–1545. doi:10.1016/S0031-9422(00)88933-5

    Article  CAS  Google Scholar 

  19. Tanaka H, Shoyama Y (1998) Formation of a monoclonal antibody against glycyrrhizin and development of an ELISA. Biol Pharm Bull 21:1391–1393

    PubMed  CAS  Google Scholar 

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  1. School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan

    Tomoyo Hasuda, Yukio Hitotsuyanagi & Koichi Takeya

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  1. Tomoyo Hasuda
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  2. Yukio Hitotsuyanagi
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  3. Koichi Takeya
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Correspondence to Koichi Takeya.

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Hasuda, T., Hitotsuyanagi, Y. & Takeya, K. Production of monoclonal antibodies against antitumor cyclohexapeptide RA-VII from Rubia cordifolia and their characterization. J Nat Med 65, 588–593 (2011). https://doi.org/10.1007/s11418-011-0508-z

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  • DOI: https://doi.org/10.1007/s11418-011-0508-z

Keywords

  • Rubia cordifolia
  • Cyclic hexapeptide
  • Monoclonal antibody
  • Cross-reactivity