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A monoclonal antibody‐based indirect competitive enzyme‐linked immunosorbent assay to quantify swertiamarin and related compounds in Swertia japonica Makino
Author(s) -
Nuntawong Poomraphie,
Horikawa Taiki,
Ochi Akihiro,
Wada Shinji,
Tsuneura Yumi,
Tanaka Hiroyuki,
Sakamoto Seiichi,
Morimoto Satoshi
Publication year - 2020
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2999
Subject(s) - chemistry , japonica , glycoside , monoclonal antibody , chromatography , high performance liquid chromatography , repeatability , traditional medicine , quantitative analysis (chemistry) , antibody , botany , stereochemistry , immunology , medicine , biology
Swertia japonica Makino ( S. japonica ) has a long history of use as a folk medicine, and it is one of the three essential Japanese folk medicines. S. japonica has been reported to have various biological activities. The biologically active secoiridoid glycoside swertiamarin (SM) has been isolated from S. japonica . The efficacy of this plant is attributed to SM and related secoiridoid glycosides. To control the quality of S. japonica for medicinal use, a method for the determination of SM and other secoiridoid glycosides in the plant is needed. Objective To produce an anti‐SM monoclonal antibody (MAb) and develop an indirect competitive enzyme‐linked immunosorbent assay (icELISA) for S. japonica standardisation and quality control. Methodology SM was conjugated to cationised bovine serum albumin (cBSA), and the SM–cBSA conjugate was used to immunise BALB/c mice. Splenocytes from the immunised mice were then fused with SP2/0 myeloma cells to produce hybridoma cells that expressed anti‐SM MAb. Results The developed icELISA was sufficiently sensitive and had a quantitative range of 0.78 to 12.5 μg/mL. Coefficients of variation below 10% indicated good repeatability. Recoveries in a spike and recovery assay ranged from 91.84% to 115.50%, which confirmed that the icELISA was accurate. The SM content measured using the icELISA was in agreement with the results of a high‐performance liquid chromatography‐ultraviolet (HPLC‐UV) assay. Conclusion The icELISA is suitable for the high‐throughput analysis of SM and other secoiridoid glycosides in S. japonica . The method is fast, economical, and reliable for S. japonica quality control.