Cyproheptadine hydrochloride (CYP), used as human and veterinary drug, has been used illegally as feed additive for food-producing animals, which could remain in food and jeopardize human health. There is a need for on-site detection of CYP residue in animal-derived food. In this study, a hapten was designed, and a specific monoclonal antibody (mAb) was developed to detect CYP with an IC₅₀ of 1.38 ng/mL and negligible cross-reactivity (CR) for other analogs. Forthermore, a high sensitive immunochromatographic assay (QBs-ICA) was developed using quantum dot nanobeads as reporters. The assay showed the linear detection range (IC₂₀-IC₈₀) of 0.03-0.52 ng/mL, the limit of detection (LOD) and visual detection limit (VDL) reached to 0.01 and 0.625 ng/mL, respectively. Spiked recovery study in pig urine and pork confirmed that the QBs-ICA was applicable for on-site testing. This assay showed better sensitivity and speedy than the reported instrumental analysis and immunoassays.

Cry toxins have been widely used in genetically modified organisms for pest control, raising public concern regarding their effects on the natural environment and food safety. In this work, a phage-mediated competitive chemiluminescent immunoassay (c-CLIA) was developed for determination of Cry1Ab toxin using anti-idiotypic camel nanobodies. By extracting RNA from camels' peripheral blood lymphocytes, a naive phage-displayed nanobody library was established. Using anti-Cry1Ab toxin monoclonal antibodies (mAbs) against the library for anti-idiotypic antibody screening, four anti-idiotypic nanobodies were selected and confirmed to be specific for anti-Cry1Ab mAb binding. Thereafter, a c-CLIA was developed for detection of Cry1Ab toxin based on anti-idiotypic camel nanobodies and employed for sample testing. The results revealed a half-inhibition concentration of developed assay to be 42.68 ± 2.54 ng/mL, in the linear range of 10.49-307.1 ng/mL. The established method is highly specific for Cry1Ab recognition, with negligible cross-reactivity for other Cry toxins. For spiked cereal samples, the recoveries of Cry1Ab toxin ranged from 77.4% to 127%, with coefficient of variation of less than 9%. This study demonstrated that the competitive format based on phage-displayed anti-idiotypic nanobodies can provide an alternative strategy for Cry toxin detection.

In this study, a designed hapten possessing the classic structure of PDE-5 inhibitors was synthesized. A monoclonal antibody (mAb) with broad recognition for six PDE-5 inhibitors was further produced. For the determination of lodenafil, methisosildenafil, mirodenafil, udenafil and tadalafil, the limit of detection (LOD) and IC₅₀ ranged from 1.01 to 26.91 ng mL^-1 and 12.75 to 278 ng mL^-1, respectively. Thereafter, a quantum dot bead-based lateral flow immunoassay (QB-LFIA) was developed, which improved the LOD and IC₅₀ to 0.32-6.52 ng mL^-1 and 7.45-133.8 ng mL^-1, respectively. Method validation was conducted using honey and capsule samples spiked with PDE-5 inhibitors, and the recoveries of the intra- and inter-assays ranged from 81.01% to 108.16%, with coefficients of variation below 12.71%. In addition, the validity and the consistency have been confirmed with a comparison between QB-LFIA and HPLC-MS/MS (R² = 0.9957). Furthermore, the developed QB-LFIA was employed for the inspection of real products, and several samples were found to be adulterated with lodenafil and methisosildenafil.