Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
47, Number 6, July 2009, pp. 467-472
Optimization of poly(GMA-co-EDMA) Monolithic
Support for Trypsin Nanoreactor Fabrication
Alex Monzo1,2, Tomas Rejtar2,
and András
Guttman1, 1Horváth Laboratory of Bioseparation Sciences, University
of Innsbruck, Austria; 2Barnett Institute of Chemical and Biological
Analysis, Northeastern University, Boston, MA
Fabrication of poly(glycidyl methacrylate-co-ethylene
dimethacrylate) [also referred to as poly(GMA-co-EDMA)] monoliths
was optimized as supporting material for trypsin digestion nanoreactors.
Reaction parameters, such as polymerization time, porogen concentration,
and monomer to crosslinker ratios, were evaluated in respect
to the permeability of the resulting monolith and their effect
on digestion efficiency, estimated by mass spectrometric analysis
of a model protein cytochrome C. The structural homogeneity of
the resulting monolithic support was checked by scanning electron
microscopy. The best nanoreactor performance, measured by the
reduction of nanoreactor backpressure and increased sequence
coverage of cytochrome C, was achieved with 8% 2-octanol (porogen)
20%/20% glycidyl methacrylate to ethylene glycol dimethacrylate
ratio and 5 h of polymerization time. Digestion of as low as
3 µg of cytochrome C with 77% sequence coverage was obtained
using the optimized trypsin nanoreactor.
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