UNIGE document Scientific Article
previous document  unige:85603  next document
add to browser collection
Title

Potential of hydrophilic interaction chromatography for the analyticalcharacterization of protein biopharmaceuticals

Authors
Beck, Alain
Lauber, Matthew
Published in Journal of Chromatography. A. 2016, vol. 1448, p. 81-92
Abstract A new stationary phase based on wide-pore hybrid silica bonded with amide ligand has been used toexplore the utility of HILIC for the analytical characterization of protein biopharmaceuticals. Various,highly-relevant samples were tested, including different insulins, interferon a-2b and trastuzumab. Thiswork shows that HILIC can be successfully employed for the analysis of therapeutic proteins and mAbs,using mobile phase compositions comprised of between 65 and 80% ACN and 0.1% TFA. In terms of elutionorder and selectivity, these HILIC separations have proven to be highly orthogonal to RPLC, while thekinetic performance remains comparable. In the case of characterizing trastuzumab, HILIC was uniquelyable to resolve several important glycoforms at the middle-up level of analysis (fragments of 25–100 kDa).Such a separation of glycoforms has been elusive by other separation mechanisms, such as RPLC and IEX.Besides showing orthogonality to RPLC and improved separations of glycoforms, HILIC offers severaladditional benefits for biopharmaceutical characterization: i) an inherent compatibility with MS, ii) areduced requirement for very high mobile phase temperatures that are otherwise needed in RPLC tolimit undesirably strong adsorption to the surface of the stationary phase, and iii) the possibility tocouple several columns in series to improve resolving power, thanks to comparatively low mobile phase viscosity.
Keywords Hydrophilic interaction chromatographyHILICInsulinsInterferon a-2bTrastuzumaba
Identifiers
Full text
Article (Published version) (2.1 MB) - document accessible for UNIGE members only Limited access to UNIGE
Structures
Research group Sciences analytiques
Citation
(ISO format)
PERIAT, Aurélie Claudine et al. Potential of hydrophilic interaction chromatography for the analyticalcharacterization of protein biopharmaceuticals. In: Journal of Chromatography. A, 2016, vol. 1448, p. 81-92. https://archive-ouverte.unige.ch/unige:85603

56 hits

0 download

Update

Deposited on : 2016-08-02

Export document
Format :
Citation style :