The objective of the current study is to analyze numerically the effect of the temperature-jump boundary condition on heterogeneous microfluidic immunosensors under electrothermal force. A three-dimensional simulation using the finite element method on the binding reaction kinetics of C-reactive protein (CRP) was performed. The kinetic reaction rate was calculated with coupled Laplace, Navier−Stokes, energy, and mass diffusion equations. Two types of reaction surfaces were studied: one in the form of a disc surrounded by two electrodes and the other in the form of a circular ring, one electrode is located inside the ring and the other outside. The numerical results reveal that the performance of a microfluidic biosensor is enhanced by using the second design of the sensing area (circular ring) coupled with the electrothermal force. The improvement factor under the applied ac field 15 Vrms was about 1.2 for the first geometry and 3.6 for the second geometry. Furthermore, the effect of temperature jump on heat transfer rise and response time was studied. The effect of two crucial parameters, viz. Knudsen number (Kn) and thermal accommodation coefficient (σT) with and without electrothermal effect, were analyzed for the two configurations.