Importance of membrane selection in the development of immunochromatographic assays for low-molecular weight compounds

This study was performed to demonstrate the importance of selecting an appropriate membrane when developing immunochromatographic assays (ICAs) for the sensitive detection of low-molecular weight compounds. Based on our findings, we propose a theoretical basis for selecting such a membrane. When eluting the sample solution for the competitive ICA using colloidal gold label for low-molecular analytes, the degree of binding inhibition is proportional to the collision frequency between the antibody-colloidal gold (Ab–CG) and analyte before Ab–CG binding to the capture antigen and a higher concentration of pesticides around the Ab–CG leads to a greater degree of inhibition. Therefore, we propose that the relative migration speed of the analyte and Ab–CG on the test strip is critically important for selecting a membrane in the development of sensitive competitive ICAs. We developed a novel method to estimate such a relative migration speed. We demonstrated the applicability of this proposal by using it to select an appropriate membrane for the development of an ICA of the pesticide diazinon.