title: Multi-Site N-glycan mapping study 1: Capillary electrophoresis - laser induced fluorescence.
abstract: 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-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra high-performance 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.
authors.affiliation: a Horváth Laboratory of Bioseparation Sciences; University of Debrecen ; Debrecen , H-4032 , Hungary.
authors.forename: Ákos
authors.initials: Á
authors.lastname: Szekrényes
authors.affiliation: b Drug Product Development; P&PD; Amgen; Inc. ; Thousand Oaks , CA 91320 , USA.
authors.forename: SungAe Suhr
authors.initials: SS
authors.lastname: Park
authors.affiliation: c SCIEX ; Brea , CA 92821 , USA.
authors.forename: Marcia
authors.initials: M
authors.lastname: Santos
authors.affiliation: c SCIEX ; Brea , CA 92821 , USA.
authors.forename: Clarence
authors.initials: C
authors.lastname: Lew
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Aled
authors.initials: A
authors.lastname: Jones
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Ted
authors.initials: T
authors.lastname: Haxo
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Kimzey
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Shiva
authors.initials: S
authors.lastname: Pourkaveh
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Szabó
authors.affiliation: e Analytical Development; Biogen ; Cambridge , MA 02142 , USA.
authors.forename: Zoran
authors.initials: Z
authors.lastname: Sosic
authors.affiliation: e Analytical Development; Biogen ; Cambridge , MA 02142 , USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Feng
authors.affiliation: a Horváth Laboratory of Bioseparation Sciences; University of Debrecen ; Debrecen , H-4032 , Hungary.
authors.forename: Csaba
authors.initials: C
authors.lastname: Váradi
authors.affiliation: f Analis s.a. R&D Diag. ; Suarlée ( Namur ), Belgium.
authors.forename: François
authors.initials: F
authors.lastname: de l'Escaille
authors.affiliation: f Analis s.a. R&D Diag. ; Suarlée ( Namur ), Belgium.
authors.forename: Jean-Bernard
authors.initials: JB
authors.lastname: Falmagne
authors.affiliation: g Bioanalytical and Discovery Analytical Sciences; Bristol-Myers Squibb ; Lawrenceville , NJ 08648 , USA.|t Current affiliation: Process Analytical, AbbVie , North Chicago , IL 60064 , USA .
authors.forename: Preeti
authors.initials: P
authors.lastname: Sejwal
authors.affiliation: h Protein Analytical Chemistry Department ; Genentech; Inc. ; South San Francisco , CA 94080 , USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Niedringhaus
authors.affiliation: h Protein Analytical Chemistry Department ; Genentech; Inc. ; South San Francisco , CA 94080 , USA.
authors.forename: David
authors.initials: D
authors.lastname: Michels
authors.affiliation: i Bioanalytical Sciences; Eli Lilly and Company (previously ImClone) ; Branchburg , NJ 08876 , USA.
authors.forename: Gordon
authors.initials: G
authors.lastname: Freckleton
authors.affiliation: j Vaccine Bioprocess Research and Development; Merck Research Laboratories ; West Point , PA 19486 , USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Hamm
authors.affiliation: k Analytical Research and Development; Pfizer; Inc. ; Andover , MA 01810 , USA.
authors.forename: Anastasiya
authors.initials: A
authors.lastname: Manuilov
authors.affiliation: l Analytical Science; Boehringer Ingelheim; Inc. ; Fremont , CA 94555 , USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Schwartz
authors.affiliation: m Regeneron Pharmaceuticals; Inc. ; Tarrytown , NY 10591 , USA.
authors.forename: Jiann-Kae
authors.initials: JK
authors.lastname: Luo
authors.affiliation: n Analytical Sciences; Seattle Genetics; Inc. ; Bothell , WA 98021 , USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: van Dyck
authors.affiliation: o ImmunoGen; Inc. ; Waltham , MA 02451 , USA.
authors.forename: Pui-King
authors.initials: PK
authors.lastname: Leung
authors.affiliation: p Analytical Research and Development; Gedeon Richter; Plc. ; Budapest , H-1475 , Hungary.
authors.forename: Marcell
authors.initials: M
authors.lastname: Olajos
authors.affiliation: q Eli Lilly and Company ; Indianapolis ; IN 46285 , USA.
authors.forename: Yingmei
authors.initials: Y
authors.lastname: Gu
authors.affiliation: r Division of Monoclonal Antibody; National Institutes for Food and Drug Control ; Beijing , PR China.
authors.forename: Kai
authors.initials: K
authors.lastname: Gao
authors.affiliation: r Division of Monoclonal Antibody; National Institutes for Food and Drug Control ; Beijing , PR China.
authors.forename: Wenbo
authors.initials: W
authors.lastname: Wang
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Jo
authors.initials: J
authors.lastname: Wegstein
authors.affiliation: d ProZyme; Inc. ; Hayward , CA 94545 , USA.
authors.forename: Samnang
authors.initials: S
authors.lastname: Tep
authors.affiliation: a Horváth Laboratory of Bioseparation Sciences; University of Debrecen ; Debrecen , H-4032 , Hungary.|s MTA-PE Translational Glycomics Group; MUKKI; University of Pannonia ; Veszprém , Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Guttman
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019075:Electrophoresis, Capillary; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry*
chemical_list: D011134:Polysaccharides
keywords: Biotherapeutics; Capillary electrophoresis; Intercompany study; N-glycans