Insulator-based dielectrophoresis (iDEP) is known as a powerful technique for separation and manipulation of bioparticles. In recent years, iDEP designs using arrays of insulating posts have shown promising results towards reaching high-efficient bioparticles manipulation. However, there is still an essential need for providing comprehensive design guidelines and further optimizing such devices. In this research, we utilized numerical simulation to study, in detail, insulating posts iDEP technique with the specific application of bioparticles separation. To achieve this, we first developed a robust numerical model to predict the electric and fluid flow fields’ distribution, and how bioparticles are being manipulated inside the system. This enabled us to study the fundamental principles of such an iDEP method. In the next step, different design aspects of insulating posts iDEP were investigated. Specifically, we focused on the effect of posts geometry and configuration on the systems’ key operation criteria such as the effectiveness of the electric field non-uniformity, the flow velocity distribution and shear stress rates. Furthermore, we studied how different electrodes’ setup may affect the electric field distribution and consequently the device performance. Finally, the developed numerical tool was used to demonstrate separation of circulating tumor cells (CTCs) from white blood cells (WBCs). For this purpose, MDA-231 breast cancer cells and Granulocytes were chosen as an indicator of CTCs ad WBCs. Our developed numerical model and presented results lay the groundwork for design and fabrication of high-efficient insulating posts iDEP microchips.