Development and characterisation of highly specific monoclonal antibody-based immunoassays for the detection and quantification of genistein-7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside in Derris scandens (Roxb.) Benth.

Autor: Sae-Foo W; Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand., Singkham S; Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand., Srisongkhram P; Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand., Yusakul G; School of Pharmacy, Walailak University, Nakhon Si Thammarat, Thailand., Masugarut P; Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand., Putalun W; Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.
Jazyk: angličtina
Zdroj: Phytochemical analysis : PCA [Phytochem Anal] 2024 Apr; Vol. 35 (3), pp. 483-492. Date of Electronic Publication: 2023 Nov 15.
DOI: 10.1002/pca.3305
Abstrakt: Introduction: The stem of the plant species Derris scandens (Roxb.) Benth. (DS) contains genistein-7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside (GTG), which is a unique marker. Previous analyses of GTG using antibody-based immunoassays were compromised because of their high cross-reactivity with structurally related compounds of DS, thereby limiting their applicability in DS quality control.
Objective: Conjugation of GTG with carrier proteins was achieved using the Mannich reaction to produce a highly specific monoclonal antibody (mAb) targeting GTG (anti-GTG mAb).
Methods: The anti-GTG mAb was generated using hybridoma technology and characterised using an indirect competitive enzyme-linked immunosorbent assay (icELISA). Both lateral-flow immunoassay (LFIA) and icELISA were developed to detect and quantify GTG in DS raw materials and associated products.
Results: icELISA using the anti-GTG mAb showed 100% specificity for GTG, with only 1.77% cross-reactivity with genistin and less than 0.01% cross-reactivity with other compounds. icELISA demonstrated a linear range for GTG determination between 62.5 and 2000 ng/mL. The limits of detection (LOD) and quantification were 49.68 and 62.50 ng/mL for GTG, respectively. The precision of the analysis ranged from 1.28% to 4.20% for repeatability and from 1.03% to 7.05% for reproducibility. The accuracy of the analysis ranged from 101.97% to 104.01% for GTG recovery. GTG levels determined via icELISA were consistent with those confirmed via high-performance liquid chromatography (HPLC) (R 2  = 0.9903). Moreover, the LOD of LFIA for GTG was 500 ng/mL.
Conclusion: Immunoassays utilising specific anti-GTG mAbs were successfully developed, including LFIA for rapid GTG detection and icELISA for GTG quantification.
(© 2023 John Wiley & Sons, Ltd.)
Databáze: MEDLINE
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