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Immobilized monolithic enzymatic reactor and its application for analysis of in‐vitro fertilization media samples
Author(s) -
Chen WeiQiang,
Obermayr Philipp,
Černigoj Urh,
Vidič Jana,
PanićJanković Tanta,
Mitulović Goran
Publication year - 2017
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201700197
Subject(s) - digestion (alchemy) , chromatography , enzymatic hydrolysis , chemistry , sample preparation , reproducibility , hydrolysis , proteomics , immobilized enzyme , enzyme , biochemistry , gene
Classical proteomics approaches involve enzymatic hydrolysis of proteins (either separated by polyacrylamide gels or in solution) followed by peptide identification using LC‐MS/MS analysis. This method requires normally more than 16 h to complete. In the case of clinical analysis, it is of the utmost importance to provide fast and reproducible analysis with minimal manual sample handling. Herein we report the method development for online protein digestion on immobilized monolithic enzymatic reactors (IMER) to accelerate protein digestion, reduce manual sample handling, and provide reproducibility to the digestion process in clinical laboratory. An integrated online digestion and separation method using monolithic immobilized enzymatic reactor was developed and applied to digestion and separation of in‐vitro‐fertilization media.