Detection of enantiomeric impurities in a simple membrane-based optical immunosensor

Currently available methods for the detection of enantiomeric impurities generally require expensive and sophisticated instrumentation. Here, we describe a simple and inexpensive membrane-based chiral immunosensor that allows quantitative determination of chiral analytes up to an enantiomer excess of 99.9%. The experimental setup is based on a competitive reaction between the analyte and a biotin-derivatized analog for the binding sites of a stereoselective antibody, which is immobilized onto a membrane. The antibody-bound analog is detected with peroxidase-conjugated avidin that converts a colorless substrate into an insoluble dye on the membrane surface. The color intensity, which is inversely related to the concentration of analyte in a sample, can be evaluated with standard image analysis programs.