Ultrasensitive antibody production strategy based on hapten property for simultaneous immunoassay

A high-quality antibody production strategy is significant for immunoassay. In this work, four general haptens were proposed based on the 3D structure and surface electrostatic potential of molecular modeling. It was found that the sensitivity and specificity of polyclonal antibodies (pAbs) mainly depended on the bond angle of shapes liked "V" between haptens and proteins and hydrophobic parts of haptens. The quantified process was employed to obtain pAbs against cyhalofop-butyl and its metabolites (CAFs), with the IC₅₀ value of 4.9 μg·L^-1 under optimal conditions. The limit of quantization (LOQ) of the ultrasensitive icELISA in brown rice was 2 μg·kg^-1. The recoveries were 74%-110%, with a coefficient of variation (CV) less than 10%. This study indicated that the hapten property approach led to an improved immunoassay.