Reactive polymer surface platform of Pentafluorophenyl Acrylate for the application of biomolecule immobilization with enhanced sensitivity

Abstract

Reactive polymer was used in the various fields such as biotechnology, photonics, and optoelectronics with advanced properties such as fluorescence, stimuli-responsiveness, and biocompatibility. Especially, bio application was developed rapidly during the past decades. Reactive polymer was utilized for the biomolecule immobilization so that the polymer act as the carrier. N-hydroxysuccinimide (NHS) is a famous representative active polymer in order to use the biomolecule conjugation. However, it is reported that NHS ester-containing polymer is not dissolves well in the organic solvent except for the DMSO, DMF. So, NHS functional group has poor solubility in most organic solvent. As alternative reactive polymer, poly pentafluorophenyl acrylate (PPFPA) as active esters have been studied a lot with very high activity. Compared to NHS, it has been studied that it has better control over polymer synthesis and has excellent stability. Also, PPFPA is possible to do post-polymerization modification (PPM) without catalyst, so that the PFP group on the PPFPA was dissociate when the amine or alcohol group is encountered into the PPFPA. After the PPM, amide or another ester bond was formed after the dissociation of the reactive pendent group. Reactive polymer with functional group was easily synthesized by CRP (controlled radical polymerization) such as NMP, ATRP and RAFT as well as post-modified to change the construction from initial structure. Among the CRP technique, RAFT is fully organic, and there’s no need toxic catalyst for the reaction, there is no need to do additional work to remove the catalyst after polymerization in this dissertation, RAFT polymerization was adopted for the PPFPA synthesis. And PPFPA after the RAFT was post-modified to immobilize the biomolecule as well as change the polymer property from hydrophobicity to hydrophilicity by using the hydrophilic molecules such as amine-PEG. Biomolecule was immobilized on the Si wafer, glass wafer and Si particle and utilized in the immunoprecipitation and detecting the virus in this dissertation. We simply introduce the dissertation in chapter 1. Synthesis of functional polymer and PPFPA, PPFPMA polymerization by RAFT was introduced in chapter 2. Also, biomolecules immobilization with reactive polymer was introduced. In chapter 3, we present the protein purification poly (pentafluorophenyl acrylate) based platform functionalized with SiO$_2$ Beads. Hepatitis virus (HCV, HAV) detection with reactive polymer platform targeting dsRNA was investigated in chapter 4. In chapter 5, we reported the multi-layered PEG-g-PPFPA polymer film based with biosensor enhanced fluorescent signal.