Preparation of a Highly Specific Single Chain Variable Fragment Antibody Targeting Miroestrol and Its Application in Quality Control of Pueraria Candollei by Enzyme-Linked Immunosorbent Assay

In 2019, a study was conducted to develop a sensitive and specific enzyme-linked immunosorbent assay (ELISA) for the quantification of miroestrol, a phytoestrogen from Pueraria candollei, using a single chain variable fragment (scFv) antibody. The scFv antibody was expressed in E. coli and used to establish an ELISA with a quantification range of 0.06–7.81 µg/mL and a limit of quantification of 0.06 µg/mL. The assay demonstrated high specificity, with cross-reactivities of less than 0.06% with related compounds, and high precision, with intra- and inter-assay relative standard deviations below 6.0%. Accuracy was validated with recovery rates between 95.08% and 103.99%. The scFv-based ELISA correlated well with a monoclonal antibody-based ELISA, with a coefficient of determination (R²) of 0.9995. The study concluded that the scFv-based ELISA is a reliable, cost-effective, and eco-friendly method for quality control of Pueraria candollei materials and supplements.