Application of sample displacement batch chromatography for fractionation of proteoforms

Abstract: Fractionation of proteoforms is currently the most challenging topic in the field of proteoform analysis. The need for considering the existence of proteoforms in experimental approaches is not only important in Life Science research in general but especially in the manufacturing of therapeutic proteins (TPs) like recombinant therapeutic antibodies (mAbs). Some of the proteoforms of TPs have significantly decreased actions or even cause side effects. The identification and removal of proteoforms differing from the main species, having the desired action, is challenging because the difference in the composition of atoms is often very small and their concentration in comparison to the main proteoform can be low. In this study, we demonstrate that sample displacement batch chromatography (SDBC) is an easy‐to‐handle, economical, and efficient method for fractionating proteoforms. As a model sample a commercial ovalbumin fraction was used, containing many ovalbumin proteoforms. The most promising parameters for the SDBC were determined by a screening approach and applied for a 10‐segment fractionation of ovalbumin with cation exchange chromatography resins. Mass spectrometry of intact proteoforms was used for characterizing the SDBC fractionation process. By SDBC, a significant separation of different proteoforms was obtained.

Standort
Deutsche Nationalbibliothek Frankfurt am Main
Umfang
Online-Ressource
Sprache
Englisch

Erschienen in
Application of sample displacement batch chromatography for fractionation of proteoforms ; day:26 ; month:09 ; year:2023 ; extent:12
Proteomics ; (26.09.2023) (gesamt 12)

Urheber
Hidayah, Siti Nurul
Biabani, Ali
Gaikwad, Manasi
Nissen, Paula
Voß, Hannah
Riedner, Maria
Schlüter, Hartmut
Siebels, Bente

DOI
10.1002/pmic.202200424
URN
urn:nbn:de:101:1-2023092615320858890431
Rechteinformation
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Letzte Aktualisierung
14.08.2025, 10:48 MESZ

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Beteiligte

  • Hidayah, Siti Nurul
  • Biabani, Ali
  • Gaikwad, Manasi
  • Nissen, Paula
  • Voß, Hannah
  • Riedner, Maria
  • Schlüter, Hartmut
  • Siebels, Bente

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