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001-es BibID:	BIBFORM084931
Első szerző:	Szekrényes Ákos (vegyészmérnök)
Cím:	Multi-Site N-glycan mapping study 1: Capillary electrophoresis ? laser induced fluorescence / Szekrényes Ákos, Park SungAe Suhr, Santos Marcia, Lew Clarence, Jones Aled, Haxo Ted, Kimzey Michael, Pourkaveh Shiva, Szabó Zoltán, Sosic Zoran, Feng Peng, Váradi Csaba, de l'Escaille François, Falmagne Jean-Bernard, Sejwal Preeti, Niedringhaus Thomas, Michels David, Freckleton Gordon, Hamm Melissa, Manuilov Anastasiya, Schwartz Melissa, Luo Jiann-Kae, van Dyck Jonathan, Leung Pui-King, Olajos Marcell, Gu Yingmei, Gao Kai, Wang Wenbo, Wegstein Jo, Tep Samnang, Guttman András
Dátum:	2016
ISSN:	1942-0862 1942-0870
Megjegyzések:	An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra highperformance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk Biotherapeutics Capillary electrophoresis Intercompany study N-glycans
Megjelenés:	mAbs. - 8 : 1 (2016), p. 56-64. -
További szerzők:	Park, SungAe Suhr Santos, Marcia Lew, Clarence Jones, Aled Haxo, Ted Kimzey, Michael Pourkaveh, Shiva Szabó Zoltán Sosic, Zoran Feng, Peng Váradi Csaba (1988-) (molekuláris biológus) de l'Escaille, François Falmagne, Jean-Bernard Sejwal, Preeti Niedringhaus, Thomas Michels, David Freckleton, Gordon Hamm, Melissa Manuilov, Anastasiya Schwartz, Melissa Luo, Jiann-Kae van Dyck, Jonathan Leung, Pui-King Olajos Marcell Gu, Yingmei Gao, Kai Wang, Wenbo Wegstein, Jo Tep, Samnang Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:	K 116263 NKFIH
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM084845
Első szerző:	Szigeti Márton (környezetmérnök)
Cím:	Fully Automated Sample Preparation for Ultrafast N-Glycosylation Analysis of Antibody Therapeutics / Szigeti Marton, Lew Clarence, Roby Keith, Guttman Andras
Dátum:	2016
ISSN:	2211-0682 1540-2452
Megjegyzések:	There is a growing demand in the biopharmaceutical industry for high-throughput, large-scale N-glycosylation profiling of therapeutic antibodies in all phases of product development, but especially during clone selection when hundreds of samples should be analyzed in a short period of time to assure their glycosylation-based biological activity. Our group has recently developed a magnetic bead?based protocol for N-glycosylation analysis of glycoproteins to alleviate the hard-to-automate centrifugation and vacuum?centrifugation steps of the currently used protocols. Glycan release, fluorophore labeling, and cleanup were all optimized, resulting in a <4h magnetic bead?based process with excellent yield and good repeatability. This article demonstrates the next level of this work by automating all steps of the optimized magnetic bead?based protocol from endoglycosidase digestion, through fluorophore labeling and cleanup with high-throughput sample processing in 96-well plate format, using an automated laboratory workstation. Capillary electrophoresis analysis of the fluorophore-labeled glycans was also optimized for rapid (<3 min) separation to accommodate the high-throughput processing of the automated sample preparation workflow. Ultrafast N-glycosylation analyses of several commercially relevant antibody therapeutics are also shown and compared to their biosimilar counterparts, addressing the biological significance of the differences.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk N-glycosylation antibody therapeutics magnetic beads automation capillary electrophoresis
Megjelenés:	Journal of Laboratory Automation. - 21 : 2 (2016), p. 281-286. -
További szerzők:	Lew, Clarence Roby, Keith Guttman András (1954-) (vegyészmérnök)
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM058340
035-os BibID:	(WoS)000337643500017 (Scopus)84902789927
Első szerző:	Váradi Csaba (molekuláris biológus)
Cím:	Rapid Magnetic Bead Based Sample Preparation for Automated and High Throughput N-Glycan Analysis of Therapeutic Antibodies / Csaba Váradi, Clarence Lew, András Guttman
Dátum:	2014
ISSN:	0003-2700
Megjegyzések:	Full automation to enable high throughput N-glycosylation profiling and sequencing with good reproducibility is vital to fulfill the contemporary needs of the biopharmaceutical industry and requirements of national regulatory agencies. The most prevalently used glycoanalytical methods of capillary electrophoresis and hydrophilic interaction liquid chromatography, while very efficient, both necessitate extensive sample preparation and cleanup, including glycoprotein capture, N-glycan release, fluorescent derivatization, purification, and preconcentration steps during the process. Currently used protocols to fulfill these tasks require multiple centrifugation and vacuum-centrifugation steps, making liquid handling robot mediated automated sample preparation difficult and expensive. In this paper we report on a rapid magnetic bead based sample preparation approach that enables full automation including all the process phases just in a couple of hours without requiring any centrifugation and/or vacuum centrifugation steps. This novel protocol has been compared to conventional glycan sample preparation strategies using standard glycoproteins (IgG, fetuin, and RNase B) and featured rapid processing time, high release and labeling efficiency, good reproducibility, and the potential of easy automation.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk glycan preparation
Megjelenés:	Analytical Chemistry. - 86 : 12 (2014), p. 5682-5687. -
További szerzők:	Lew, Clarence Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:	MTA-PE Translation Glycomics project MTA
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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