Application of sample displacement batch chromatography for fractionation of proteoforms
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Application of sample displacement batch chromatography for fractionation of proteoforms. / Hidayah, Siti Nurul; Biabani, Ali; Gaikwad, Manasi; Nissen, Paula; Voß, Hannah; Riedner, Maria; Schlüter, Hartmut; Siebels, Bente.
In: PROTEOMICS, Vol. 24, No. 3-4, e2200424, 02.2024.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Application of sample displacement batch chromatography for fractionation of proteoforms
AU - Hidayah, Siti Nurul
AU - Biabani, Ali
AU - Gaikwad, Manasi
AU - Nissen, Paula
AU - Voß, Hannah
AU - Riedner, Maria
AU - Schlüter, Hartmut
AU - Siebels, Bente
N1 - © 2023 The Authors. Proteomics published by Wiley-VCH GmbH.
PY - 2024/2
Y1 - 2024/2
N2 - 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.
AB - 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.
U2 - 10.1002/pmic.202200424
DO - 10.1002/pmic.202200424
M3 - SCORING: Journal article
C2 - 37750450
VL - 24
JO - PROTEOMICS
JF - PROTEOMICS
SN - 1615-9853
IS - 3-4
M1 - e2200424
ER -
