Sample preparation for N-glycosylation analysis of therapeutic
monoclonal antibodies by electrophoresis

C 2015

Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis

SZEKRÉNYES, Ákos; Jan PARTYKA; Csaba VARADI; Jana KRENKOVA; František FORET et. al.

Základní údaje

Originální název

Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis

Autoři

SZEKRÉNYES, Ákos (348 Maďarsko); Jan PARTYKA (203 Česká republika, domácí); Csaba VARADI (348 Maďarsko); Jana KRENKOVA (203 Česká republika); František FORET (203 Česká republika, garant, domácí) a András GUTTMAN (348 Maďarsko)

Vydání

NEW YORK, Methods in Molecular Biology, od s. 183-195, 13 s. Microchip Capillary Electrophoresis Protocols, 1274, 2015

Nakladatel

Springer Science+Business Media

Další údaje

Jazyk

angličtina

Typ výsledku

Kapitola resp. kapitoly v odborné knize

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Forma vydání

tištěná verze "print"

Odkazy

URL

Kód RIV

RIV/00216224:14740/15:00082381

Organizační jednotka

Středoevropský technologický institut

ISBN

978-1-4939-2352-6

DOI

http://dx.doi.org/10.1007/978-1-4939-2353-3_16

EID Scopus

2-s2.0-84922879525

Klíčová slova anglicky

capillary electrophoresis; monoclonal antibodies; sample preparation; Fluorophore labeling; N-glycan analysis; Therapeutic monoclonal antibody

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 29. 4. 2016 12:48, Mgr. Eva Špillingová

Anotace

V originále

There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans.

Citovat

SZEKRÉNYES, Ákos; Jan PARTYKA; Csaba VARADI; Jana KRENKOVA; František FORET a András GUTTMAN. Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. In Ann Van Schepdael. Methods in Molecular Biology. NEW YORK: Springer Science+Business Media, 2015, s. 183-195. Microchip Capillary Electrophoresis Protocols, 1274. ISBN 978-1-4939-2352-6. Dostupné z: https://dx.doi.org/10.1007/978-1-4939-2353-3_16.

@inbook{1226563,
   author = {Szekrényes, Ákos and Partyka, Jan and Varadi, Csaba and Krenkova, Jana and Foret, František and Guttman, András},
   address = {NEW YORK},
   booktitle = {Methods in Molecular Biology},
   doi = {http://dx.doi.org/10.1007/978-1-4939-2353-3_16},
   editor = {Ann Van Schepdael},
   keywords = {capillary electrophoresis; monoclonal antibodies; sample preparation; Fluorophore labeling; N-glycan analysis; Therapeutic monoclonal antibody},
   howpublished = {tištěná verze "print"},
   language = {eng},
   location = {NEW YORK},
   isbn = {978-1-4939-2352-6},
   pages = {183-195},
   publisher = {Springer Science+Business Media},
   title = {Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis},
   url = {https://link.springer.com/protocol/10.1007%2F978-1-4939-2353-3_16},
   year = {2015}
}

TY  - CHAP
ID  - 1226563
AU  - Szekrényes, Ákos - Partyka, Jan - Varadi, Csaba - Krenkova, Jana - Foret, František - Guttman, András
PY  - 2015
TI  - Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis
VL  - Microchip Capillary Electrophoresis Protocols, 1274
PB  - Springer Science+Business Media
CY  - NEW YORK
SN  - 9781493923526
KW  - capillary electrophoresis
KW  - monoclonal antibodies
KW  - sample preparation
KW  - Fluorophore labeling
KW  - N-glycan analysis
KW  - Therapeutic monoclonal antibody
UR  - https://link.springer.com/protocol/10.1007%2F978-1-4939-2353-3_16
L2  - https://link.springer.com/protocol/10.1007%2F978-1-4939-2353-3_16
N2  - There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans.
ER  -

SZEKRÉNYES, Ákos; Jan PARTYKA; Csaba VARADI; Jana KRENKOVA; František FORET a András GUTTMAN. Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. In Ann Van Schepdael. \textit{Methods in Molecular Biology}. NEW YORK: Springer Science+Business Media, 2015, s.~183-195. Microchip Capillary Electrophoresis Protocols, 1274. ISBN~978-1-4939-2352-6. Dostupné z: https://dx.doi.org/10.1007/978-1-4939-2353-3\_{}16.

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