Details

Autor(en) / Beteiligte

• Engel, Nicole Y.
• Puffler, Nicole
• Marchetti-Deschmann, Martina
• Allmaier, Günter
• Weiss, Victor U.

Titel

nES-DMA with Charge-reduction based on Soft X-ray Radiation: Analysis of a Recombinant Monoclonal Antibody

Ist Teil von

• Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2021-10, Vol.1182, p.122925-122925, Article 122925

Ort / Verlag

Elsevier B.V

Erscheinungsjahr

2021

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• •Nano differential mobility analysis applied in characterization of a recombinant monoclonal antibody (rmAb)•Intact rmAb and aggregates detectable in a native-like environment.•rmAb fragments after IdeS protease digestion detectable next to intact rmAb.•Molecular weights derived from gas-phase electrophoresis match MS data.•rmAb production monitoring and final quality control possible. Due to the fast growing importance of monoclonal antibodies in biomedical research, bioanalytics and human therapy, sensitive, fast and reliable methods are needed to monitor their production, target their characteristics, and for their final quality control. Application of a nano electrospray (nES) with soft X-ray radiation (SXR) based charge reduction and differential mobility analysis (DMA, aka nano electrospray gas-phase electrophoretic mobility molecular analysis, nES GEMMA) allows the size-separation and detection of macromolecules and (bio-)nanoparticles from a few nm up to several hundreds of nm in diameter in a native-like environment. The current study focuses on the analysis of a 148 kDa recombinant monoclonal antibody (rmAb) with the above mentioned instrumental setup and applying an universal detector, i.e. a water-based condensation particle detector (CPC). Next to the intact rmAb, its aggregates and fragment products after digestion with IdeS protease were analyzed. Additionally, influence of temperature treatment and pH variation on the stability of the rmAb was monitored. In this context, changes in electrophoretic mobility diameter (EMD) values, peak shape, and signal intensity based on particle numbers were of interest. Molecular weights calculated by application of a correlation derived from respective standard protein compounds were compared to mass spectrometric values and were found to be in good accordance. To conclude, we demonstrate that nES-DMA is a valuable tool in the characterization and quality control of rmABs.