Biomolecule activated surfaces have become a versatile material platform with applications in areas such as delivery, separation, and detection. In particular, reactive pentafluorophenyl acrylate (PFPA) polymers have been used in the preparation of biomolecule immobilized surfaces due to the high reactivity of pentafluorophenyl (PFP) groups towards amine-containing molecules. In this contribution, we report the preparation and application of poly(PFPA) based platform for detection of viral double-stranded RNAs (dsRNAs). Long dsRNAs are signature of virus, and they are detected in cells infected with single-stranded RNA virus, dsRNA virus, and positive-strand single-stranded DNA virus. Mammalian cells have developed innate immune response that recognizes long stretch of dsRNA to induce immune response. Inspired by this immune mechanism, we utilize dsRNA recognizing J2 antibody and reactive poly(PFPA) to develop a simple method to quickly capture and detect dsRNA. Poly(PFPA) is synthesized via RAFT polymerization then grafted to solid substrate using 3-aminopropyltriethoxysilane (APTES) as a linker molecule. Utilizing the exchange reaction between the PFP units on the polymer and the amine groups on J2, the antibodies are then immobilized with high efficiency. The antibody immobilized surface is then successfully used to detect fluorescent tagged synthetic dsRNAs. Furthermore, to achieve label-free target recognition, a two-stage dsRNA detection scheme is developed where fluorescent tagged antibodies are used in the visualization of the bound dsRNAs. This study shows the development and application of J2 antibody immobilized poly(PFPA) as a diagnostic platform for dsRNA based viral infection.

Reactive polymer-based dsRNA detection as a diagnostic platform for viral infection

2019 Fall National Meeting

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