Termed as the third generation of flow injection analysis, the lab-on-valve (LOV) system is closely related to the principle of sequential injection analysis and constructed aiming at facilitating various miniature analytical operations and downscaling sample/reagent consumption. The highly integrated configuration makes LOV system compatible with virtually all kinds of spectroscopic detection techniques and presents it an intelligent platform for bead-injection protocol. The minimized reagent consumption and precise fluidic manipulation also propose the LOV system an excellent candidate for the adaptation of sample processing in the macroscopic world with the microfluidic analysis system. After a few years development, it has been applied extensively in the miniaturization and automation of analytical procedures, especially in the field of bioassays. This review presents the up-to-date progress and some of the selected applications of the LOV system for bioanalytical assays. In addition, future perspectives are discussed.