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Application of sample displacement batch chromatography for fractionation of proteoforms
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  • Siti Nurul Hidayah,
  • Manasi Gaikwad,
  • Ali Biabani,
  • Paula Nissen,
  • Bente Siebels,
  • Hannah Voß,
  • Maria Riedner,
  • Hartmut Schlüter
Siti Nurul Hidayah
University Medical Center Hamburg-Eppendorf
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Manasi Gaikwad
University Medical Center Hamburg-Eppendorf
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Ali Biabani
University Medical Center Hamburg-Eppendorf
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Paula Nissen
University Medical Center Hamburg-Eppendorf
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Bente Siebels
University Medical Center Hamburg-Eppendorf
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Hannah Voß
University Medical Center Hamburg-Eppendorf
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Maria Riedner
University of Hamburg
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Hartmut Schlüter
University Medical Center Hamburg-Eppendorf

Corresponding Author:hschluet@uke.de

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Abstract

Fractionation of proteoforms is currently the most challenging topic in the field of protein purification. The need for considering the existence of proteoforms into 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 often is very small and their concentration in comparison to the main proteoform can be small. In this study we demonstrate that sample displacement batch chromatography (SDBC) is an easy to handle, economic 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 the ovalbumin with cation exchange chromatography resin. Mass spectrometry of intact proteoforms was used for characterizing the SDBC fractionation process. By SDBC a significant separation of different proteoforms was obtained.
08 Mar 2023Submitted to PROTEOMICS
09 Mar 2023Submission Checks Completed
09 Mar 2023Assigned to Editor
09 Mar 2023Review(s) Completed, Editorial Evaluation Pending
09 Mar 2023Reviewer(s) Assigned
23 Mar 2023Editorial Decision: Revise Minor
28 Jul 2023Review(s) Completed, Editorial Evaluation Pending
28 Jul 20231st Revision Received
09 Aug 2023Reviewer(s) Assigned
11 Aug 2023Editorial Decision: Revise Minor
06 Sep 2023Review(s) Completed, Editorial Evaluation Pending
06 Sep 20232nd Revision Received
07 Sep 2023Editorial Decision: Accept