The working principle of immunoassays is based on the specific binding reaction of an analyte-ligand protein pair in physiological environments. However, for a diffusion-limited protein, the diffusion boundary layer of the analyte on the reaction surface of a biosensor would hinder the binding reaction from association and dissociation. The formation of such association and dissociation layers thus limits the response time and the overall performance of a biosensor. In this work we have performed a two-dimensional full time scale finite element simulation on the binding reaction kinetics of two commonly used proteins, C-reactive protein ͑CRP͒ and immunoglobulin G ͑IgG͒. By applying a nonuniform ac electric field to the flow microchannel of the biosensor, the electrothermal force can be generated to induce a pair of vortices to stir the flow field. With the aid of the vortices and a suitable choice of the location of the biosensor, the fluids flowing over the reacting surface can be accelerated fast enough to depress efficiently the growth of the diffusion boundary layer on the reaction surface, and enhance the association or dissociation of analyte-ligand complex. The interference patterns of the flow field due to the existence of the sensor at different locations of the microchannel could cause different degrees of enhancement to the association and the dissociation. By changing the location of the sensor the largest enhancement is found at the position near the negative electrode. For the configuration of the microchannel we studied, the initial slope of the curve of the analyte-ligand complex versus time can be raised up to 5.17 for CRP and 1.93 for IgG in association, and 3.74 for CRP and 1.28 for IgG in dissociation, respectively, under the applied ac field 15 V rms peak-to-peak and operating frequency 100 kHz. At this optimal sensor location, we also studied the effect of various settings of temperature boundary conditions on the top and bottom walls, including the two limiting cases, namely, constant temperature and thermal insulation on both walls. We show that varying the temperature boundary conditions can cause an essential effect on the enhancement of the binding reaction and can be employed to find an optimal binding enhancement. Utilizing these simulation results, an improved design incorporating a pair of electrodes and a neck region near the reaction surface is demonstrated. The sensor is fixed to locate at the middle of the bottom side. With the existence of the stirring flow field, the association rate of the 30 m neck is 2.73 times faster than that of the original channel with no neck.
Biochemical applications of microchips often require a rapid mixing of different fluid samples. At the microscale level, fluid flow is usually a highly ordered laminar flow and diffusion is the primary mechanism for mixing owing to the lack of disturbances, yielding inefficiency for practical biochemical analysis. In this work, we design a prototype active micromixer by employing the electrothermal effect. We apply to the flow microchannel a non-uniform AC electric field, which can generate an electrothermal force on the fluid flow and induce vortex pairs for enhancing mixing efficiency. The performance of this active micromixer is studied and compared, under the same mixing quality, with that of a conventional passive micromixer of the same size with obstacles in the flow channel by three-dimensional finite element simulations. The numerical results show that the pressure drop between the inlet and the outlet for the active micromixer is much less than (only 3000th) that for the passive micro-mixer with the same mixing quality. To obtain an optimal mixing quality, we have systematically studied the mixing quality by varying the geometrical arrangements of the electrodes. An almost complete mixing can be obtained using a specific design. Moreover, the temperature increases around the electrodes are lower than 3 K, which does not adversely affect the biochemical analysis. It is suggested that the prototype active micromixer designed is promising and effective and useful for biochemical analysis.
We investigate a immunoassay biosensor that employs a Quartz Crystal Microbalance (QCM) to detect the specific binding reaction of the (Human IgG1)-(Anti-Human IgG1) protein pair under physiological conditions. In addition to experiments, a three dimensional time domain finite element method (FEM) was used to perform simulations for the biomolecular binding reaction in microfluidic channels. In particular, we discuss the unsteady convective diffusion in the transportation tube, which conveys the buffer solution containing the analyte molecules into the micro-channel where the QCM sensor lies. It is found that the distribution of the analyte concentration in the tube is strongly affected by the flow field, yielding large discrepancies between the simulations and experimental results. Our analysis shows that the conventional assumption of the analyte concentration in the inlet of the micro-channel being uniform and constant in time is inadequate. In addition, we also show that the commonly used procedure in kinetic analysis for estimating binding rate constants from the experimental data would underestimate these rate constants due to neglected diffusion processes from the inlet to the reaction surface. A calibration procedure is proposed to supplement the basic kinetic analysis, thus yielding better consistency with experiments.
After the Code Case N-640 was issued in 1999, the fracture toughness curve of reactor pressure vessel materials in ASME Section XI-Appendix G was amended to the KIC curve. In Taiwan, the present pressure-temperature limit curves of normal reactor startup (heat-up) and shut-down (cool-down) for the reactor pressure vessel is still calculated per KIA curve in 1998 or earlier editions. In this paper, the failure risks of a Taiwan domestic reactor pressure vessel under various pressure-temperature limit operations are analyzed. First, the pressure-temperature limit curves of the Taiwan domestic reactor pressure vessel based on KIA and KIC curves, and various levels of embrittlement, are calculated. Then, the ORNL’s probabilistic fracture mechanics code, FAVOR, and the PNNL’s flaw model are utilized to assess the failure probabilities of the reactor pressure vessel under such pressure-temperature limit transients. Further, the deterministic analyses of FAVOR code are also conducted. It is found that under the pressure-temperature limit transients based on KIC curves, the reactor pressure vessel presents higher failure probabilities, but are all below the allowable risk. The present results indicate that using the KIC curve the pressure-temperature limits can either increase the operational margin or still maintains the sufficient stability of the analyzed reactor pressure vessel.
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