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Publication
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| Title of Article |
Development and characterisation of highly specificmonoclonal antibody-based immunoassays for the detectionand quantification of genistein-7-O-[α-rhamnopyranosyl-(1!6)]-β-glucopyranoside inDerris scandens(Roxb.) Benth. |
| Date of Acceptance |
30 October 2023 |
| Journal |
| Title of Journal |
Phytochemical Analysis |
| Standard |
SCOPUS |
| Institute of Journal |
John Wiley & Sons, Ltd. |
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| Year of Publication |
2023 |
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| Abstract |
Introduction:The stem of the plant speciesDerris 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 immunoas-says were compromised because of their high cross-reactivity with structurallyrelated compounds of DS, thereby limiting their applicability in DS quality control.Objective:Conjugation of GTG with carrier proteins was achieved using the Mannichreaction to produce a highly specific monoclonal antibody (mAb) targeting GTG (anti-GTG mAb).Methods:The anti-GTG mAb was generated using hybridoma technology and char-acterised using an indirect competitive enzyme-linked immunosorbent assay (icE-LISA). Both lateral-flow immunoassay (LFIA) and icELISA were developed to detectand quantify GTG in DS raw materials and associated products.Results:icELISA using the anti-GTG mAb showed 100% specificity for GTG, withonly 1.77% cross-reactivity with genistin and less than 0.01% cross-reactivity withother compounds. icELISA demonstrated a linear range for GTG determinationbetween 62.5 and 2000 ng/mL. The limits of detection (LOD) and quantificationwere 49.68 and 62.50 ng/mL for GTG, respectively. The precision of the analysis ran-ged from 1.28% to 4.20% for repeatability and from 1.03% to 7.05% for reproducibil-ity. 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 ofLFIA for GTG was 500 ng/mL.Conclusion:Immunoassays utilising specific anti-GTG mAbs were successfully devel-oped, including LFIA for rapid GTG detection and icELISA for GTG quantification. |
| Keyword |
Derris scandens, ELISA, lateral-flow immunoassay, Mannich reaction, monoclonal antibody |
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| Reviewing Status |
มีผู้ประเมินอิสระ |
| Status |
ตีพิมพ์แล้ว |
| Level of Publication |
นานาชาติ |
| citation |
false |
| Part of thesis |
true |
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| Citation |
0
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Journal Publication
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