Download PDF
Planta Med 2007; 73(10): 1127-1132
DOI: 10.1055/s-2007-981560
Analytical Methods
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Preparation of a Novel Monoclonal Antibody against the Antimalarial Drugs, Artemisinin and Artesunate

Hiroyuki Tanaka1 , Waraporn Putalun2 , Wanchai De-Eknamkul3 , Oraphan Matangkasombut4 , Yukihiro Shoyama1
  • 1Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
  • 2Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
  • 3Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand
  • 4Faculty of Pharmacy, Rangsit University, Pathumthani, Thailand
Further Information

Publication History

Received: March 2, 2007 Revised: May 30, 2007

Accepted: June 4, 2007

Publication Date:
05 July 2007 (online)

Also available at
    Permissions and Reprints

Abstract

A monoclonal antibody (MAb) 1C1 against artemisinin was prepared by a cell fusion with splenocytes and aminopterin-sensitive myeloma cells, SP2/0. An artesunate-BSA conjugate was used as immunogen to raise antibodies specific to artemisinin. The prepared anti-artemisinin MAb-1C1 have a novel characteristic which shows a specificity to compounds that are structurally related to artemisinin such as artesunate (630 %) and dihydroartemisinin (29.9 %). By using MAb-1C1, a specific and reliable ELISA was developed for the detection of compounds structurally related to artemisinin. The system shows a full measuring range from 2 to 20 μg/mL in the case of artemisinin and from 4 to 125 ng/mL in the case of artesunate in the competitive ELISA, and was validated to be of use for surveying and breeding of Artemisia annua containing a high amount of artemisinin as well as for the characterization of the pharmacokinetics of artemisinin and its derivatives.

Key words

Artemisinin - artesunate - Artemisia annua - Asteraceae - ELISA - monoclonal antibody

References

  • 1 Phillips R S. Current status of malaria and potential for control.  Clin Microb Reviews. 2001;  14 208-26.
    PubMedGoogle Scholar
  • 2 Zheng G Q. Cytotoxic terpenoids and flavonoids from Artemisia annua .  Planta Med. 1994;  60 54-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Beekman A C, Wierenga P K, Woerdenbag H J, Van Uden W, Pras N, Konings A W. et al . Artemisinin-derived sesquiterpene lactones as potential antitumour compounds: cytotoxic action against bone marrow and tomour cells.  Planta Med. 1998;  64 615-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Romeo M R, Serrano M A, Vallejo M, Efferth T, Alvarez M, Marin J J. Antiviral effect of artemisinin from Artemisia annua against a model member of the Flaviviridae family, the bovine viral diarrhea virus (BVDV).  Planta Med. 2006;  72 1169-74.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Efferth T. Willmar Schwabe Award 2006: Antiplasmodial and antitumor activity of artemisinin-from bench to bedside.  Planta Med. 2007;  73 299-309.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Zhao S, Zeng M Y. Application of precolumn reaction to high performance liquid chromatography of Qinghausu in animal plasma.  Anal Chem. 1986;  58 289-92.
    CrossrefPubMedGoogle Scholar
  • 7 Elsohly H N, Croom E M, Elsohly M A. Analysis of the antimalarial sesquiterpene artemisinin in Artemisia annua by high-performance liquid-chromatography (HPLC) with postcolumn derivatization and ultraviolet detection.  Pharm Res. 1987;  4 258-60.
    CrossrefPubMedGoogle Scholar
  • 8 Batty K T, Davis T ME, Thu L A, Binh T Q, Anh T K, Ilett K F. Selective high-performance liquid chromatographic determination of artesunate and alpha- and beta-dihydroartemisinin in patients with falciparum malaria.  J Chromatogr B. 1996;  677 345-50.
    PubMedGoogle Scholar
  • 9 Jastrebova J, Nyholm L, Markides K E, Bergqvist Y. On-line deoxygenation for reductive electrochemical detection of artemisinin and dihydroartemisinin in liquid chromatography.  Analyst. 1998;  123 313-7.
    CrossrefPubMedGoogle Scholar
  • 10 Karbwang J, NaBangchang K, Molunto P, Banmairuroi V, Congpuong K. Determination of artemether and its major metabolite, dihydroartemisinin, in plasma using high-performance liquid chromatography with electrochemical detection.  J Chromatogr B. 1997;  690 259-65.
    PubMedGoogle Scholar
  • 11 Melendez V, Peggins J O, Brewer T G, Theoharides A D. Determination of the antimalarial arteether and its deethyated metabolite dehydroartemisinin in plasma by high-performance liquid-chromatography with reductive electrochemical detection.  J Pharm Sci. 1991;  80 132-8.
    CrossrefPubMedGoogle Scholar
  • 12 Peys E, Vandenkerckhove J, Van Hemel J, Sas B. Simultaneous determination of beta-artemether and its metabolite dihydroartemisinin in human plasma and urine by a high-performance liquid chromatography-mass spectrometry assay using electrospray ionization.  Chromatographia. 2005;  61 637-41.
    CrossrefPubMedGoogle Scholar
  • 13 Wang M F, Park C H, Wu Q L, Simon J E. Analysis of artemisinin in Artemisia annua L. by LC-MS with selected ion monitoring.  J Agric Food Chem. 2005;  53 7010-3.
    CrossrefPubMedGoogle Scholar
  • 14 Rajanikanth M, Madhusudanan K P, Gupta R C. An HPLC-MS method for simultaneous estimation of alpha,beta-arteether and its metabolite dihydroartemisinin, in rat plasma for application to pharmacokinetic study.  Biomed Chromatogr. 2003;  17 440-6.
    CrossrefPubMedGoogle Scholar
  • 15 Peng C A, Ferreira J FS, Wood A J. Direct analysis of artemisinin from Artemisia annua L. using high-performance liquid chromatography with evaporative light scattering detector, and gas chromatography with flame ionization detector.  J Chromatogr A. 2006;  1133 254-8.
    CrossrefPubMedGoogle Scholar
  • 16 Liu C Z, Zhou H Y, Zhao Y. An effective method for fast determination of artemisinin in Artemisia annua L. by high performance liquid chromatography with evaporative light scattering detection.  Anal Chin Acta. 2007;  581 298-302.
    PubMedGoogle Scholar
  • 17 Sakata R, Shoyama Y, Murakami H. Production of monoclonal-antibodies and enzyme-linked immunoassay for typical adenylate-cyclase activator, forskolin.  Cytotechnology. 1994;  16 101-8.
    CrossrefPubMedGoogle Scholar
  • 18 Tanaka H, Goto Y, Shoyama Y. Monoclonal antibody based enzyme immunoassay for marihuana (cannabinoid) compounds.  J Immunoassay. 1996;  17 321-42.
    PubMedGoogle Scholar
  • 19 Xuan L J, Tanaka H, Xu Y M, Shoyama Y. Preparation of monoclonal antibody against crocin and its characterization.  Cytotechnology. 1999;  29 65-70.
    CrossrefPubMedGoogle Scholar
  • 20 Tanaka H, Fukuda N, Shoyama Y. Formation of monoclonal antibody against a major ginseng component, ginsenoside Rb1 and its characterization.  Cytotechnology. 1999;  29 115-20.
    CrossrefPubMedGoogle Scholar
  • 21 Morinaga O, Tanaka H, Shoyama Y. Production of monoclonal antibody against a major purgative component, sennoside A, its characterization and ELISA.  Analyst. 2000;  125 1109-13.
    CrossrefPubMedGoogle Scholar
  • 22 Shan S J, Tanaka H, Shoyama Y. Enzyme-linked immunosorbent assay for glycyrrhizin using anti-glycyrrhizin monoclonal antibody and an eastern blotting technique for glucuronides of glycyrrhetic acid.  Anal Chem. 2001;  3 5784-90.
    PubMedGoogle Scholar
  • 23 Kim J S, Tanaka H, Shoyama Y. Immunoquantitative analysis for berberine and its related compounds using monoclonal antibodies in herbal medicines.  Analyst. 2004;  129 87-91.
    CrossrefPubMedGoogle Scholar
  • 24 Goto Y, Shima Y, Morimoto S, Shoyama Y, Murakami H, Kusai A. et al . Determination of tetrahydrocannabinolic acid-carrier protein conjugate by matrix-assisted laser-desorption ionization mass-spectrometry and antibody-formation.  Org Mass Spectrom. 1994;  29 668-71.
    CrossrefPubMedGoogle Scholar
  • 25 Galfre G, Milstein C. Preparation of monoclonal antibodies: strategies and procedures.  Methods Enzymol. 1981;  73 3-46.
    PubMedGoogle Scholar
  • 26 Goding J W. Antibody-production by hybridomas.  J Immunol Methods. 1980;  39 285-308.
    CrossrefPubMedGoogle Scholar
  • 27 Murakami H, Masui H, Sato G H, Sueoka N, Chow T P, Kanosueoka T. Growth of hybridoma cells in serum-free medium-ethanolamine is an essential component.  Proc Natl Acad Sci USA. 1982;  79 1158-62.
    CrossrefPubMedGoogle Scholar
  • 28 Weiler E W, Zenk M H. Radioimmunoassay for determination of digoxin and related compounds in Digitalis lanata .  Phytochemistry. 1976;  15 1537-45.
    CrossrefPubMedGoogle Scholar

Hiroyuki Tanaka

Department of Pharmacognosy

Graduate School of Pharmaceutical Sciences

Kyushu University

3-1-1 Maidashi

Higashi-ku

Fukuoka 812-8582

Japan

Email: htanaka@phar.kyushu-u.ac.jp