Optimization of poly(GMA-co-EDMA) monolithic support for trypsin nanoreactor fabrication

Alex Monzo, Tomas Rejtar, A. Guttman

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)467-472
Number of pages6
JournalJournal of Chromatographic Science
Volume47
Issue number6
Publication statusPublished - Jul 2009

Fingerprint

Nanoreactors
Trypsin
Cytochromes
Fabrication
Polymerization
Monomers
Scanning electron microscopy
GMA-EDMA
Proteins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Optimization of poly(GMA-co-EDMA) monolithic support for trypsin nanoreactor fabrication. / Monzo, Alex; Rejtar, Tomas; Guttman, A.

In: Journal of Chromatographic Science, Vol. 47, No. 6, 07.2009, p. 467-472.

Research output: Contribution to journalArticle

@article{07033fc9623e467287c74ea97cdf4b9c,
title = "Optimization of poly(GMA-co-EDMA) monolithic support for trypsin nanoreactor fabrication",
abstract = "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.",
author = "Alex Monzo and Tomas Rejtar and A. Guttman",
year = "2009",
month = "7",
language = "English",
volume = "47",
pages = "467--472",
journal = "Journal of Chromatographic Science",
issn = "0021-9665",
publisher = "Preston Publications",
number = "6",

}

TY - JOUR

T1 - Optimization of poly(GMA-co-EDMA) monolithic support for trypsin nanoreactor fabrication

AU - Monzo, Alex

AU - Rejtar, Tomas

AU - Guttman, A.

PY - 2009/7

Y1 - 2009/7

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=68349109777&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68349109777&partnerID=8YFLogxK

M3 - Article

C2 - 19555552

AN - SCOPUS:68349109777

VL - 47

SP - 467

EP - 472

JO - Journal of Chromatographic Science

JF - Journal of Chromatographic Science

SN - 0021-9665

IS - 6

ER -