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Development and characterisation of highly specific monoclonal antibody-based immunoassays for the detection and quantification of genistein-7-O-[α-rhamnopyranosyl-(1→6)]-ß-glucopyranoside in Derris scandens (Roxb.) Benth.
Sae-Foo, Worapol; Singkham, Sukritta; Srisongkhram, Pimsiri; Yusakul, Gorawit; Masugarut, Pisitchai; Putalun, Waraporn.
Afiliación
  • 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.
Phytochem Anal ; 35(3): 483-492, 2024 Apr.
Article en En | MEDLINE | ID: mdl-37965872
INTRODUCTION: The stem of the plant species Derris scandens (Roxb.) Benth. (DS) contains genistein-7-O-[α-rhamnopyranosyl-(1→6)]-ß-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) (R2 = 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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Derris / Anticuerpos Monoclonales Idioma: En Revista: Phytochem Anal Asunto de la revista: BOTANICA / QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Tailandia Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Derris / Anticuerpos Monoclonales Idioma: En Revista: Phytochem Anal Asunto de la revista: BOTANICA / QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Tailandia Pais de publicación: Reino Unido