SIP-HTM based noro-virus sensor using virus-like particles

Abstract

Surface imprinted polymers (SIP) are biomimetic receptors that have active binding cavities on their surface, and that selectively bind particles similar with template particles used in the production process. SIPs can be synthesized in a few ways, and since 2001 SIPs have been used in many applications. When a SIP is coupled with a detection platform, it can act as a biosensor revealing the presence of small particles such as viruses. SIPs have been coupled with a quartz crystal microbalance (QCM), impedance spectroscopy, surface plasmon resonance and thermal methods. In the group of Prof. Patrick Wagner at KU Leuven, a heat transfer method (HTM) is being developed that observes the change in thermal conduction due to binding of the particles on the surface of the polymer.

Virus-like particles (VLP) are particles that resemble viruses but are not infectious since they do not contain the genetic material inside. Vesa Hytönens group at Tampere University has been synthesizing Noro-VLPs in insect cells using the baculovirus expression system. The ultimate goal of the collaboration project is to produce SIP-HTMs using -VLPs as template particles, to create a biosensor that could be used to detect human viruses from liquid samples. In this Thesis work, some of the necessary fabrication steps were explored. Viruses have not been detected before using SIP-HTM and such detectors could be invaluable tools in controlling epidemics and monitoring the environment, for example with food safety issues.

In this thesis I introduce general methods used for virus detection and also the SIP-HTM sensor that is tried to be fabricated in this project. The adsorption of the VLP-particles on a PDMS surface is a vital part of the fabrication of the SIP. I have experimentally studied the adsorption of VLPs on different surfaces with varying properties. A few protocols were found following which the VLP adsorption could be optimized. Using AFM and HIM, extensive imaging of VLPs and SIPs has been performed during this fabrication step. Images showed surface structure and topology of SIPs made using VLPs and yeasts as template particles.