Original title: Oriented immobilization of PNgase F on a porous polymer monolith for rapid N-glycan release
Authors: Szekrényes, A. ; Křenková, Jana ; Keresztessy, Z. ; Foret, František ; Guttman, A.
Document type: Papers
Conference/Event: CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./, Brno (CZ), 2012-11-01 / 2012-11-02
Year: 2012
Language: eng
Abstract: 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.
Keywords: monolith; oriented immobilization; PNGase F
Project no.: 2SGA2721
Funding provider: Jihomoravský kraj
Host item entry: CECE 2012. 9th International Interdisciplinary Meeting on Bioanalysis, ISBN 978-80-904959-1-3
Note: Související webová stránka: http://hdl.handle.net/11104/0215253

Institution: Institute of Analytical Chemistry AS ČR (web)
Document availability information: Fulltext is available in the digital repository of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0215253

Permalink: http://www.nusl.cz/ntk/nusl-135527


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