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Immobilized endoproteinase G lu‐ C to magnetic bead cellulose as a tool in proteomic analysis
Author(s) -
Prikryl Petr,
Ticha Marie,
Kucerova Zdenka
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300118
Subject(s) - cellulose , bead , chemistry , chromatography , magnetic bead , biochemistry , materials science , composite material
Magnetic bead cellulose activated with divinyl sulfone was used for the immobilization of S taphylococcus aureus endoproteinase G lu‐ C (EC 3.4.21.19). The immobilized proteinase was characterized by increased thermostability, by decreased self‐cleavage activity, and a possibility of repeated use. The prepared immobilized enzyme was applied for the proteolytic cleavage of α‐casein and BSA under different conditions (different composition of buffers, different pH, and different time of digestion). The possibilities of the direct use of enzyme reaction products for MALDI TOF MS analysis were shown.