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Reversed-Phase Liquid Chromatography for the Analysis of Therapeutic Proteins and Recombinant Monoclonal Antibodies

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Published in LCGC Europe. 2012, vol. 25, no. 10, p. 540-550
Abstract Some promising reversed-phase widepore stationary phases (organic monolithic, fully porous and core-shell type) were recently introduced for the fast and high resolution separations of large biomolecules. It was demonstrated that kinetic perfomance, retention capability, selectivity for closely related proteins and loading capacity, as well as recoveries, were satisfactory with recent core-shell and sub-2µm fully porous materials. This article focuses on the achievable throughput and resolution for the characterization of biomolecules. As shown, fast separation of a therapeutic protein (~19 kDa) and some of its degradants (related proteins) was achieved in less than 1.5 min with a 50-mm column length while a peak capacity of more than 150 was obtained in about 15 min using two columns of 150 mm coupled in series. Similar results were also attained with intact therapeutic monoclonal antibodes (mAbs) of ~150 kDa and their fragments by varying the column length and gradient time. However, the recovery of intact mAbs was found to be unacceptable in revered-phase liquid chromatography (RPLC) except when using elevated mobile phase temperature (80-90°C).
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FEKETE, Szabolcs, GUILLARME, Davy. Reversed-Phase Liquid Chromatography for the Analysis of Therapeutic Proteins and Recombinant Monoclonal Antibodies. In: LCGC Europe, 2012, vol. 25, n° 10, p. 540-550. https://archive-ouverte.unige.ch/unige:28344

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Deposited on : 2013-06-03

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