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Planta Med 2014; 80(04): 337-342
DOI: 10.1055/s-0033-1360394
Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Preparation of a Monoclonal Antibody against Notoginsenoside R1, a Distinctive Saponin from Panax notoginseng, and Its Application to Indirect Competitive ELISA

Supattra Limsuwanchote
1   Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai, Songkhla, Thailand
,
Juraithip Wungsintaweekul
1   Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai, Songkhla, Thailand
,
Gorawit Yusakul
3   Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Jing-Yan Han
2   Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China
,
Kaori Sasaki-Tabata
3   Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Hiroyuki Tanaka
3   Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Yukihiro Shoyama
4   Department of Pharmacognosy, Nagasaki International University, Sasebo, Nagasaki, Japan
,
Satoshi Morimoto
3   Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
› Author Affiliations
Further Information

Publication History

received 28 August 2013
revised 07 January 2014

accepted 20 January 2014

Publication Date:
07 March 2014 (online)

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Abstract

We have prepared a monoclonal antibody against notoginsenoside R1, a primary active constituent of Sanqi ginseng (roots of Panax notoginseng). The monoclonal antibody was raised by immunizing BALB/c male mice with notoginsenoside R1-bovine albumin conjugates following cell fusion via electroporation. This method has been shown to be very effective for producing hybridomas with excellent antibody prevalence following cell fusion of their splenocytes with cells of the myeloma cell line SP2/0. Of all the hybridomas secreting a monoclonal antibody against notoginsenoside R1, only the 1A1P cell line produces a highly specific antibody to this compound. Surprisingly, the cross-reactivity of this monoclonal antibody for ginsenoside Rg1 and ginsenoside Re, derivatives of protopanaxatriol, was below 1.02 % in a competitive immunoassay. Based on this characteristic of the monoclonal antibody, an indirect competitive ELISA (range of measurement 0.56–9 µg/mL) was established and applied as a quality control method. In conclusion, the developed immunoassay was easy to handle and reliable for the analysis of notoginsenoside R1 in Sanqi ginseng products without requiring a pretreatment.

Key words

notoginsenoside R1 - ELISA - monoclonal antibody - Panax notoginseng - Araliaceae - Sanqi ginseng
 

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