Virus Purification by Bead-Based Immunoreaction and Rapid Electrokinetic Patterning

Abstract

Recently bio-detection techniques are developed to carry out the concept of Lab-on-a-chip (LOC) for time saving and sample reduction Particle manipulation techniques are useful to bead-based applications Rapid electrokinetic patterning (REP) is an optoelectrokinetic tool for particle manipulation REP features low laser power addressable manipulation and free of complicated fabrication As a result we conducted virus purification based on REP to prove its capability in the field of LOC In virology virus purification is necessary because the virus concentration in the blood samples is extremely low Therefore large sample volume is required in some purification methods such as the density-gradient centrifugation However it is time consuming and it cannot separate substance with similar density Combining the bead-based immunoreaction with REP system to achieve the virus concentration The study was separated and evaluated in three steps First step: capture virus by bead-based immunoreaction; second step: sort particles by REP; third step: extract the concentrated sample by droplet separation This study investigated the feasibility of each step separately in a simulated manner The results showed the first step the capability of the virus capturing is evaluated by virus-mimicking gold nanoparticles (AuNPs) The same capability is confirmed when using Hepatitis B (HBV) and Enterovirus 71 (EV71) samples but the efficiency was interfered by virus-like-particles (VLPs) and antigens Different sizes of particles is mixed to simulate the virus captured particles sorting from the other components in a sample in the second step Sorting particles into two sides cannot be reached by applying different electric frequency but it can be achieved by moving the laser spot to increase the difference of particle assembly speed From the experiment the particles can be concentrated on one side of the droplet by REP manipulation and the separated efficiency reached 93% In the third step making hydrophilic and hydrophobic regions can be used in separating a droplet into two individual ones This study proved the feasibility of three steps process in biomedical application but according to previous research converting the purpose to protein simultaneous purification with detection would be a better strategy Furthermore sorting particle by unequal response speed of different sizes of particles has never been proposed it can be an issue of the further research on REP

Date of Award	2017 Feb 14
Original language	English
Supervisor	Han-Sheng Chuang (Supervisor)