Single-reagent Fluorescence Polarization Immunoassay for Melamine

doi:10.7503/cjcu20120346

Abstract

Abstract:

Fluorescence-labeled melamine tracer(MEL-EDF) was synthesized via the reaction of melamine hapten(MEL) with 1-(2-aminoethyl)-3-(3',6'-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-5-yl)thiourea(EDF). The tracer was then incubated with the antibody to prepare the single reagent immunocomplex, and both the binding and displacement kinetics were evaluated to obtain the equilibration time. Finally, a single reagent fluorescence polarization immunoassay(SR-FPIA) was developed to measure melamine. The SR-FPIA method demonstrated a detection limit(LOD) of 3.2 ng/mL, a 50% inhibition value(IC₅₀) of 157.1 ng/mL and a working range(IC₂₀-IC₈₀) of 12.2-1012.4 ng/mL. The results indicated the proposed method can meet the detection requirement for the maximum residue level of melamine set by Chinese government and Codex Alimentarius Commission(CAC). The detection could be finished within 15 min, and the obtained single reagent immunocomplex was stable for more than 30 days at 4℃. This work will benefit to further development of SR-FPIA kit for melamine.

Key words: Single-reagent fluorescence polarization immunoassay, Tracer, Melamine

CLC Number:

O657

TrendMD:

WANG Qiang, HUANG Qi-Xin, SUN Yuan-Ming, LIU Ying-Ju, XU Zhen-Lin, LEI Hong-Tao. Single-reagent Fluorescence Polarization Immunoassay for Melamine[J]. Chem. J. Chinese Universities, 2013, 34(1): 67.

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Single-reagent Fluorescence Polarization Immunoassay for Melamine

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