Development of an enzyme immunoassay using a monoclonal antibody against the psychoactive diterpenoid salvinorin A

Madan Kumar Paudel, Osamu Shirota, Kaori Tabata, Hiroyuki Tanaka, Setsuko Sekita, Satoshi Morimoto

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Salvinorin A (1), the main active constituent in Salvia divinorum, is a highly selective kappa-opioid receptor agonist with hallucinogenic effects, which is regulated in several countries. In the present study, a monoclonal antibody (mAb) against 1 was prepared, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) system was developed for the detection of salvinorins. To raise mAbs against 1, salvinorin B (2) hemisuccinate was synthesized and used to prepare the immunogen 2-bovine serum albumin conjugate. This technique was used to prepare a hybridoma cell line, 3D5, which secreted a mAb that recognized 1. The mAb was shown to have specificity for 1 and other salvinorins in cross-reactivity tests. The intra-assay calibration range by icELISA using the mAb against 1 was 0.0195-0.625 μg/mL. After validating the icELISA using intra- and interassays, a recovery experiment and analysis of several plants in the family Lamiaceae, including S. divinorum, confirmed that the analytical method based on ELISA is not only simple but also precise, accurate, sensitive, and sufficiently reliable. The results indicate that icELISA is a useful tool in the identification of S. divinorum.

Original languageEnglish
Pages (from-to)1654-1660
Number of pages7
JournalJournal of Natural Products
Volume76
Issue number9
DOIs
Publication statusPublished - Sep 27 2013

Fingerprint

salvinorin A
Diterpenes
Immunosorbents
Immunoenzyme Techniques
Assays
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Enzymes
Salvia
Lamiaceae
kappa Opioid Receptor
Hybridomas
Bovine Serum Albumin
Calibration
Cells
Recovery
Cell Line

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science
  • Drug Discovery
  • Complementary and alternative medicine
  • Organic Chemistry

Cite this

Development of an enzyme immunoassay using a monoclonal antibody against the psychoactive diterpenoid salvinorin A. / Paudel, Madan Kumar; Shirota, Osamu; Tabata, Kaori; Tanaka, Hiroyuki; Sekita, Setsuko; Morimoto, Satoshi.

In: Journal of Natural Products, Vol. 76, No. 9, 27.09.2013, p. 1654-1660.

Research output: Contribution to journalArticle

Paudel, Madan Kumar ; Shirota, Osamu ; Tabata, Kaori ; Tanaka, Hiroyuki ; Sekita, Setsuko ; Morimoto, Satoshi. / Development of an enzyme immunoassay using a monoclonal antibody against the psychoactive diterpenoid salvinorin A. In: Journal of Natural Products. 2013 ; Vol. 76, No. 9. pp. 1654-1660.
@article{c89b40b8e87b4d188b27694cc09ae6e8,
title = "Development of an enzyme immunoassay using a monoclonal antibody against the psychoactive diterpenoid salvinorin A",
abstract = "Salvinorin A (1), the main active constituent in Salvia divinorum, is a highly selective kappa-opioid receptor agonist with hallucinogenic effects, which is regulated in several countries. In the present study, a monoclonal antibody (mAb) against 1 was prepared, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) system was developed for the detection of salvinorins. To raise mAbs against 1, salvinorin B (2) hemisuccinate was synthesized and used to prepare the immunogen 2-bovine serum albumin conjugate. This technique was used to prepare a hybridoma cell line, 3D5, which secreted a mAb that recognized 1. The mAb was shown to have specificity for 1 and other salvinorins in cross-reactivity tests. The intra-assay calibration range by icELISA using the mAb against 1 was 0.0195-0.625 μg/mL. After validating the icELISA using intra- and interassays, a recovery experiment and analysis of several plants in the family Lamiaceae, including S. divinorum, confirmed that the analytical method based on ELISA is not only simple but also precise, accurate, sensitive, and sufficiently reliable. The results indicate that icELISA is a useful tool in the identification of S. divinorum.",
author = "Paudel, {Madan Kumar} and Osamu Shirota and Kaori Tabata and Hiroyuki Tanaka and Setsuko Sekita and Satoshi Morimoto",
year = "2013",
month = "9",
day = "27",
doi = "10.1021/np400358n",
language = "English",
volume = "76",
pages = "1654--1660",
journal = "Journal of Natural Products",
issn = "0163-3864",
publisher = "American Chemical Society",
number = "9",

}

TY - JOUR

T1 - Development of an enzyme immunoassay using a monoclonal antibody against the psychoactive diterpenoid salvinorin A

AU - Paudel, Madan Kumar

AU - Shirota, Osamu

AU - Tabata, Kaori

AU - Tanaka, Hiroyuki

AU - Sekita, Setsuko

AU - Morimoto, Satoshi

PY - 2013/9/27

Y1 - 2013/9/27

N2 - Salvinorin A (1), the main active constituent in Salvia divinorum, is a highly selective kappa-opioid receptor agonist with hallucinogenic effects, which is regulated in several countries. In the present study, a monoclonal antibody (mAb) against 1 was prepared, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) system was developed for the detection of salvinorins. To raise mAbs against 1, salvinorin B (2) hemisuccinate was synthesized and used to prepare the immunogen 2-bovine serum albumin conjugate. This technique was used to prepare a hybridoma cell line, 3D5, which secreted a mAb that recognized 1. The mAb was shown to have specificity for 1 and other salvinorins in cross-reactivity tests. The intra-assay calibration range by icELISA using the mAb against 1 was 0.0195-0.625 μg/mL. After validating the icELISA using intra- and interassays, a recovery experiment and analysis of several plants in the family Lamiaceae, including S. divinorum, confirmed that the analytical method based on ELISA is not only simple but also precise, accurate, sensitive, and sufficiently reliable. The results indicate that icELISA is a useful tool in the identification of S. divinorum.

AB - Salvinorin A (1), the main active constituent in Salvia divinorum, is a highly selective kappa-opioid receptor agonist with hallucinogenic effects, which is regulated in several countries. In the present study, a monoclonal antibody (mAb) against 1 was prepared, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) system was developed for the detection of salvinorins. To raise mAbs against 1, salvinorin B (2) hemisuccinate was synthesized and used to prepare the immunogen 2-bovine serum albumin conjugate. This technique was used to prepare a hybridoma cell line, 3D5, which secreted a mAb that recognized 1. The mAb was shown to have specificity for 1 and other salvinorins in cross-reactivity tests. The intra-assay calibration range by icELISA using the mAb against 1 was 0.0195-0.625 μg/mL. After validating the icELISA using intra- and interassays, a recovery experiment and analysis of several plants in the family Lamiaceae, including S. divinorum, confirmed that the analytical method based on ELISA is not only simple but also precise, accurate, sensitive, and sufficiently reliable. The results indicate that icELISA is a useful tool in the identification of S. divinorum.

UR - http://www.scopus.com/inward/record.url?scp=84885072501&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885072501&partnerID=8YFLogxK

U2 - 10.1021/np400358n

DO - 10.1021/np400358n

M3 - Article

C2 - 23987562

AN - SCOPUS:84885072501

VL - 76

SP - 1654

EP - 1660

JO - Journal of Natural Products

JF - Journal of Natural Products

SN - 0163-3864

IS - 9

ER -