Recombinant single-chain variable fragment-based immunoassay for the authentication of Vietnamese ginseng via quantitative detection of majonoside R2

IF 7.5 2区医学 Q1 CHEMISTRY, MEDICINAL

Journal of Ginseng Research Pub Date : 2026-03-01 Epub Date: 2026-01-06 DOI:10.1016/j.jgr.2025.100974

Jiranan Chaingam , Poomraphie Nuntawong , Le Van Huy , Sornkanok Vimolmangkang , Varalee Yodsurang , Seiichi Sakamoto

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引用次数: 0

Abstract

Background

Majonoside R2 (MR2) is an ocotillol-type saponin found at 3 %–5 % (w/w) in Panax vietnamensis that serves as a critical biomarker for authenticating sought-after Vietnamese ginseng varieties, such as Ngoc Linh and Lai Chau. However, widespread adulteration with morphologically similar species has created an urgent need for high-throughput, cost-effective authentication methods beyond chromatographic or deoxyribonucleic acid-based techniques.

Methods

A single-chain variable fragment (scFv) antibody against MR2 was recombinantly expressed in Escherichia coli, purified, and functionally validated. Using this antibody, an indirect competitive enzyme-linked immunosorbent assays (icELISA) approach was optimized for MR2 quantification in complex herbal matrices. The performance of this novel assay was benchmarked against that of a previously validated method using monoclonal antibody (mAb)-based ELISA and verified using authentic Panax spp. and Randia siamensis samples.

Results and conclusion

The scFv-based icELISA demonstrated a detection range of 31.25–1000 ng/mL for MR2, with high reproducibility and accuracy (CV <12.4 %, recovery 90.6 %–116.7 %). The assay was found to reliably distinguish authentic Ngoc Linh and Lai Chau ginsengs from other Panax species with high specificity. Recombinant expression in E. coli enables scalable, animal-free production, facilitating the broader adoption of this icELISA across laboratories equipped with standard molecular biology infrastructure. These findings support the integration of scFv-based assays into regulatory frameworks for authenticating Vietnamese ginseng.

Abstract Image

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重组单链可变片段免疫法定量检测越南人参皂苷R2。

背景：Majonoside R2 （MR2）是一种在越南人参中含量为3% - 5% （w/w）的醇型皂苷，是鉴定越南人参品种（如玉林和丽州）的重要生物标志物。然而，由于形态相似物种的广泛掺假，迫切需要高通量、高成本效益的鉴定方法，而不是色谱或基于脱氧核糖核酸的技术。方法：在大肠杆菌中重组表达抗MR2的单链可变片段（scFv）抗体，纯化并进行功能验证。使用该抗体，优化了间接竞争性酶联免疫吸附测定（icELISA）方法，用于复杂草药基质中MR2的定量。这种新方法的性能与先前使用基于单克隆抗体（mAb）的ELISA验证方法的性能进行了基准测试，并使用真实的Panax spp.和Randia siamensis样品进行了验证。结果与结论：基于scfv的icELISA对MR2的检测范围为31.25 ~ 1000 ng/mL，重现性和准确性高（CV Panax种特异性高）。在大肠杆菌中的重组表达实现了可扩展的、无动物的生产，促进了在配备标准分子生物学基础设施的实验室中更广泛地采用该icELISA。这些发现支持将基于scfv的检测方法整合到越南人参认证的监管框架中。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Ginseng Research CHEMISTRY, MEDICINAL-INTEGRATIVE & COMPLEMENTARY MEDICINE

CiteScore

11.40

自引率

9.50%

发文量

111

审稿时长

6-12 weeks

期刊介绍： Journal of Ginseng Research (JGR) is an official, open access journal of the Korean Society of Ginseng and is the only international journal publishing scholarly reports on ginseng research in the world. The journal is a bimonthly peer-reviewed publication featuring high-quality studies related to basic, pre-clinical, and clinical researches on ginseng to reflect recent progresses in ginseng research. JGR publishes papers, either experimental or theoretical, that advance our understanding of ginseng science, including plant sciences, biology, chemistry, pharmacology, toxicology, pharmacokinetics, veterinary medicine, biochemistry, manufacture, and clinical study of ginseng since 1976. It also includes the new paradigm of integrative research, covering alternative medicinal approaches. Article types considered for publication include review articles, original research articles, and brief reports. JGR helps researchers to understand mechanisms for traditional efficacy of ginseng and to put their clinical evidence together. It provides balanced information on basic science and clinical applications to researchers, manufacturers, practitioners, teachers, scholars, and medical doctors.