ISOELECTRIC FOCUSING DIFFERENTIAL SEPARATION ASSAY

摘要

A method for detecting and quantitating analytes among closely related substances by reacting the analyte in a test sample with excess labeled binding agent which specifically binds to the analyte to form a complex. The complex and excess labeled binding agent, which have different pIs, are separated by isoelectric focusing, with the location (x1) of the excess labeled binding agent defining an expected location (w1) at which the complex should be found. The focused substances are detected by optically scanning the separation medium (10) before or after release of the applied electric field. A second labeled marker with a known concentration provides a means for normalizing the peak areas which are compared with stored normalized calibration data to determine the amount of target analyte in the test sample.