This thesis presents practical applications of optoelectrofluidic platform for measurement and detection of biomolecules. Optoelectrofluidics refers to the study of the motions of particles or molecules and their interactions with surrounding fluid and electric field, which is induced or perturbed in an optical manner. Here a simple and most widely-used optoelectrofluidic platform, called optoelectrofluidic tweezers (OET), has been utilized for manipulating micro-/nanoparticles and molecules. Although many types of OET platform has been developed and many studies for programmable manipulation of various target objects using it has been reported until now, any practical applications such as measurement and detection of biological molecules has never been demonstrated. Here, in order to apply an optoelectrofluidic platform for measurement of molecular mobility and detection of human tumor markers, separation and concentration of microparticles, and dynamic control of colloidal assembly and local chemical concentration have been performed at first. In addition, the frequency-dependent behavior of micro-/nanoparticles and molecules in an optoelectrofluidic device has been investigated. On the basis of the basic studies about the optoelectrofluidic manipulation of micro-/nanoparticles and molecules, new schemes for measuring diffusion coefficient of molecules and for conducting sandwich immunoassays have been developed. The optoelectrofluidic technologies provide a simple, rapid and easy way to measure the diffusion coefficients of various dextran molecules, and to detect human tumor marker based on surface-enhanced Raman scattering. These measurement and detection technologies based on optoelectrofluidics open a new way for simple, automated, fast, accurate and precise measurement and detection of biomolecules. In addition, this thesis provides future perspectives about practical applications of optoelectrofluidics in biology and chemistry fields.