Oriented immobilization of PNgase F on a porous polymer monolith for rapid N-glycan release

0386014 - ÚIACH 2013 RIV CZ eng C - Conference Paper (international conference)
Szekrényes, A. - Křenková, Jana - Keresztessy, Z. - Foret, František - Guttman, A.
Oriented immobilization of PNgase F on a porous polymer monolith for rapid N-glycan release.
CECE 2012. 9th International Interdisciplinary Meeting on Bioanalysis. Brno: Ústav analytické chemie AV ČR, v. v. i, 2012 - (Foret, F.; Křenková, J.; Guttman, A.; Klepárník, K.; Boček, P.), s. 36-38. ISBN 978-80-904959-1-3.
[CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./. Brno (CZ), 01.11.2012-02.11.2012]
Grant - others:Jihomoravský kraj(CZ) 2SGA2721
Program: 2SGA
Institutional support: RVO:68081715
Keywords : PNGase F * oriented immobilization * monolith
Subject RIV: CB - Analytical Chemistry, Separation

We demonstrate a simple and rapid method for the oriented immobilization of peptide-N4-(Nacetyl- glucosaminyl) asparagine amidase F (PNGase F) on a porous polymer monolith. The oriented immobilization is based on the affinity of glutathione-S-transferase (GST) tagged PNGase F towards glutathione modified monolith prepared in the capillary format. This approach allows the oriented and easily replaceable immobilization of PNGase F for rapid and efficient release of N-linked glycans. The reactor was tested by deglycosylation of several native glycoproteins such as ribonuclease B, fetuin or human immunoglobulin G. The proteins were effectively deglycosylated in several minutes and the released N-linked glycans were analyzed using off-line MALDI/MS or CE/LIF.
Permanent Link: http://hdl.handle.net/11104/0215253