The present invention relates to a reliable, robust and sensitive platform aimed to analyze the massive kinetic profile of new molecules against its main target and also against other potential targets. Thus, the present invention relates to a method for calculating the kinetic profile of a compound of interest against a target protein or polyprotein wherein it is not needed to predetermine the Ki value of the compound of interest against the target protein or polyprotein before starting the assay. The present invention also discloses the use of said method in a high-throughput system for developing a Binding Kinetic Profiling assay of multiple compounds of interest against a unique target, or a Kinetic Selectivity Profiling assay of one selected compound against multiple target proteins or polyproteins to therefore establish multiple clinical profiles of potential drugs.