Matrix Supported Poly(2-oxazoline)-Based Hydrogels
for DNA Catch and Release
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Abstract
We describe the synthesis of matrix
supported hydrogel structures
based on amine containing poly(2-oxazoline)s and their use to bind
and release genetic material for potential applications in diagnostics
or pathogen detection. Amine containing poly(2-oxazoline)s were synthesized
by copolymerization of 2-ethyl-2-oxazoline with a monomer bearing
a <i>tert</i>-butyl oxycarbonyl (Boc) protected amine group
in the 2-position and subsequent deprotection. The statistical copolymers
were used to generate hydrogels and matrix supported hydrogels by
cross-linking of a certain fraction of the amine groups with epichlorhydrin.
Supported structures were prepared by soaking porous polyethylene
(PE) or polypropylene (PP) filter materials in a copolymer/epichlorhydrin
solution, which was cross-linked upon heating. Scanning electron microscopy
(SEM) of the composites revealed a bead like structure of the gel
phase, which could be attributed to a lower critical solution temperature
(LCST) behavior of the initial polymer prior to gelation. The dependency
of the LCST behavior on the content of amine groups was investigated.
Swelling values and the ratio of hydrogel per composite was determined
using water sorption analysis. Subsequently, the ability of the systems
to absorb and release labeled DNA was tested. Uptake and stimulated
release, triggered by changes in pH, temperature, and heparin concentration,
were investigated using fluorescence microscopy. Polymerase chain
reaction (PCR) proved the successful recovery of the DNA, demonstrating
the potential of the presented system for a broad range of molecular
biological applications