Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Lucie KoreckáMichael Przybylski

Abstract

The newly developed immobilized enzyme reactors (IMERs) with proteolytic enzymes chymotrypsin, trypsin or papain were used for specific fragmentation of high molecular-mass and heterogeneous glycoproteins immunoglobulin G (IgG) and crystallizable fragment of IgG (Fc). The efficiency of splitting or digestion were controlled by RP-HPLC. The specificity of digestion by trypsin reactor was controlled by MS. IMERs (trypsin immobilized on magnetic microparticles focused in a channel of magnetically active microfluidic device) was used for digestion of the whole IgG molecule. The sufficient conditions for IgG digestion in microfluidic device (flow rate, ratio S:E, pH, temperature) were optimized. It was confirmed that the combination of IMERs with microfluidic device enables efficient digestion of highly heterogeneous glycoproteins such as IgG in extremely short time and minimal reaction volume.

References

Aug 28, 2001·Journal of Chromatography. a·L RiggsF E Regnier
May 16, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Z BílkováJ Churácek
May 16, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Zuzana BílkováJaroslav Churácek

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Citations

Aug 14, 2012·Applied Microbiology and Biotechnology·Chrystelle GanachaudKrister Holmberg
Jun 28, 2012·Applied Biochemistry and Biotechnology·Petr PrikrylZdenka Kucerova
Feb 4, 2005·Journal of Separation Science·Gabriella Massolini, Enrica Calleri
Mar 9, 2012·Colloids and Surfaces. B, Biointerfaces·Sang Youn HwangE K Lee
Feb 13, 2009·Proteomics·Carolin HuhnManfred Wuhrer
Apr 12, 2013·Journal of Separation Science·Petr PrikrylZdenka Kucerova
Mar 21, 2016·Biochemical and Biophysical Research Communications·Guihua RuanFuyou Du
Aug 2, 2005·Biotechnology Advances·Pawel L UrbanNeil C Bruce
Dec 24, 2005·Lab on a Chip·Nicole Pamme
Aug 29, 2007·Analytica Chimica Acta·Guihua Eileen YueJerome P Ferrance
Aug 16, 2005·Lab on a Chip·Marcela SlovakovaJean-Louis Viovy
Jun 7, 2019·Biotechnology Journal·Nicole UlmerMassimo Morbidelli
Apr 2, 2013·Chemical Reviews·Martin GerickePedro Fardim

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