Research Institute for Chromatography & Metablys

Kortrijk, Belgium

Research Institute for Chromatography & Metablys

Kortrijk, Belgium
SEARCH FILTERS
Time filter
Source Type

Vanhoenacker G.,Research Institute for Chromatography & Metablys | Vandenheede I.,Research Institute for Chromatography & Metablys | David F.,Research Institute for Chromatography & Metablys | Sandra P.,Research Institute for Chromatography & Metablys | Sandra K.,Research Institute for Chromatography & Metablys
Analytical and Bioanalytical Chemistry | Year: 2014

Comprehensive two-dimensional liquid chromatography (LC×LC) is here proposed as a novel tool for peptide mapping of therapeutic monoclonal antibodies in both R&D and routine (QA/QC) environments. This is illustrated by the analysis of the tryptic digest of trastuzumab (Herceptin) applying a commercially available two-dimensional 2D-LC system. Three different LC×LC combinations, i.e., strong cation-exchange × reversed-phase (SCX×RP), reversed-phase × reversed-phase (RP×RP), and hydrophilic interaction × reversed-phase (HILIC×RP), are reported. Detection was carried out using both UV detection (DAD) and mass spectrometry (MS). Several challenges related to the application of LC×LC in peptide mapping and the hyphenation to MS are addressed. The applicability of LC×LC in the assessment of identity, purity, and comparability is demonstrated by the analysis of different Herceptin innovator production batches, a Herceptin biosimilar in development and of stressed samples. The described methodology was shown to be precise in terms of peak volume and 2D retention time opening interesting perspectives for use in QA/QC testing. © 2014 Springer-Verlag Berlin Heidelberg


PubMed | Research Institute for Chromatography & Metablys
Type: Evaluation Studies | Journal: Analytical and bioanalytical chemistry | Year: 2015

Comprehensive two-dimensional liquid chromatography (LCLC) is here proposed as a novel tool for peptide mapping of therapeutic monoclonal antibodies in both R&D and routine (QA/QC) environments. This is illustrated by the analysis of the tryptic digest of trastuzumab (Herceptin) applying a commercially available two-dimensional 2D-LC system. Three different LCLC combinations, i.e., strong cation-exchange reversed-phase (SCXRP), reversed-phase reversed-phase (RPRP), and hydrophilic interaction reversed-phase (HILICRP), are reported. Detection was carried out using both UV detection (DAD) and mass spectrometry (MS). Several challenges related to the application of LCLC in peptide mapping and the hyphenation to MS are addressed. The applicability of LCLC in the assessment of identity, purity, and comparability is demonstrated by the analysis of different Herceptin innovator production batches, a Herceptin biosimilar in development and of stressed samples. The described methodology was shown to be precise in terms of peak volume and (2)D retention time opening interesting perspectives for use in QA/QC testing.

Loading Research Institute for Chromatography & Metablys collaborators
Loading Research Institute for Chromatography & Metablys collaborators