High-sensitivity and rapid immunoassays for furosine detection based on monoclonal antibody 1C3

Abstract

Furosine, a hazardous compound mainly produced by the Maillard reaction in food at high temperatures, has been shown to exert cytotoxic effects on cells and cause liver damage after excessive consumption. Hence, it is desirable to develop correlative sensitive and robust immunoassays for furosine detection in real-world samples. In this study, Ovalbumin-furosine (OVA-furo) conjugates were employed as the immunogens for animal immunization, and a hybridoma (1C3) secreting an anti-furosine monoclonal antibody (mAb) was successfully screened. The purified 1C3 mAb exhibited high specificity and affinity (7.4 × 10⁸ L/mol), and an indirect competitive ELISA (ic-ELISA) based on the 1C3 mAb was developed for the detection of furosine. The linear detection range of the ic-ELISA was 6.03-817.85 ng/mL, with a limit of detection (LOD) value of 2 ng/mL. Colloid gold nanospheres(AuNP-) and nanoflower gold-particles(AuNF)-based strips were also established based on this antibody, which had qLOD values and linear ranges of 40 and 20 ng/mL and 40-140 and 20-90 ng/mL, respectively. All the immunoassays showed good detection performance, indicating that these immunoassays have the potential for practical application for the inexpensive detection of furosine in real-world samples.

Keywords: ELISA; Furosine; Gold nanoparticles; Immunochromatographic strip; Monoclonal antibody.