Universal probe with oriented antibody to improve the immunochromatographic assay of lead ions in Procambarus clarkii

Based on the information from the random inspection of foods by the China Food and Drug Administration in 2022, the contamination levels of lead ions are high in many edible products. Traditional methods of detecting lead ions cannot meet the requirements of on-site analysis of food due to the need for large equipment. The immunochromatographic assay (ICA) is an effective, rapid, on-site analytical technique for determining lead ions in foods. However, the performance of ICA based on the traditional probe (AuNP-mAb) is limited by ignoring the influence of the antibody orientation. In this study, we developed an efficient technology for constructing a universal probe (AuNP-PrA-mAb) based on oriented immobilization of antibody. The performance of ICA was largely improved due to specific binding of the Fc region of the antibody with recombinant protein A (PrA) on the surface of a gold nanoparticle (AuNP). The ICA based on a universal probe was applied for the qualitative and quantitative detection of lead ions in Procambarus clarkii within 30 min. Meanwhile, a simple and fast pretreatment method based on dilute acid extraction was developed for pretreating the Procambarus clarkii containing lead ions. The visual limit of detection (vLOD) and the scanning limit of quantization (sLOQ) of the developed ICA strip for lead ions were 0.5 ng/mL and 0.28 ng/mL, respectively. The sensitivity of ICA based on universal probe was 10-fold higher than that of the ICA using traditional probe. Furthermore, the detection results had no obvious difference between the ICA and ICP-MS with T-test statistical method. The developed ICA that is based on a universal probe presented broad application prospects in detecting contaminants in foods.