Change of charge variant composition of trastuzumab upon stressing at physiological conditions

B Spanov; O Olaleye; N Lingg; AEH Bentlage; N Govorukhina; J Hermans; de Merbel N van; G Vidarsson; A Jungbauer; R Bischoff

doi:10.1016/j.chroma.2021.462506

Change of charge variant composition of trastuzumab upon stressing at physiological conditions

B Spanov, O Olaleye, N Lingg, AEH Bentlage, N Govorukhina, J Hermans, de Merbel N van, G Vidarsson, A Jungbauer, R Bischoff

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Abstract

Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and in vivo because of the susceptibility of monoclonal antibodies to undergo chemical modifications. Modifications may adversely affect the potency of the drug, induce immunogenicity or affect pharmacokinetics. In this study, we evaluated the application of optimized pH gradient systems for the separation of charge variants of trastuzumab after forced degradation study. pH gradient-based elution resulted in high-resolution separation of some 20 charge variants after 3 weeks at 37°C under physiological conditions. The charge variants were further characterized by LC-MS-based peptide mapping. There was no significant difference in the binding properties to HER2 or a range of Fcγ receptors between non-stressed and stressed trastuzumab.

Original language	English
Article number	462506
Journal	Journal of chromatography A
Volume	1655
DOIs	https://doi.org/10.1016/j.chroma.2021.462506
Publication status	Published - 11 Oct 2021
Externally published	Yes

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title = "Change of charge variant composition of trastuzumab upon stressing at physiological conditions",

abstract = "Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and in vivo because of the susceptibility of monoclonal antibodies to undergo chemical modifications. Modifications may adversely affect the potency of the drug, induce immunogenicity or affect pharmacokinetics. In this study, we evaluated the application of optimized pH gradient systems for the separation of charge variants of trastuzumab after forced degradation study. pH gradient-based elution resulted in high-resolution separation of some 20 charge variants after 3 weeks at 37°C under physiological conditions. The charge variants were further characterized by LC-MS-based peptide mapping. There was no significant difference in the binding properties to HER2 or a range of Fcγ receptors between non-stressed and stressed trastuzumab.",

author = "B Spanov and O Olaleye and N Lingg and AEH Bentlage and N Govorukhina and J Hermans and van, {de Merbel N} and G Vidarsson and A Jungbauer and R Bischoff",

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doi = "10.1016/j.chroma.2021.462506",

language = "English",

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T1 - Change of charge variant composition of trastuzumab upon stressing at physiological conditions

AU - Spanov, B

AU - Olaleye, O

AU - Lingg, N

AU - Bentlage, AEH

AU - Govorukhina, N

AU - Hermans, J

AU - van, de Merbel N

AU - Vidarsson, G

AU - Jungbauer, A

AU - Bischoff, R

PY - 2021/10/11

Y1 - 2021/10/11

N2 - Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and in vivo because of the susceptibility of monoclonal antibodies to undergo chemical modifications. Modifications may adversely affect the potency of the drug, induce immunogenicity or affect pharmacokinetics. In this study, we evaluated the application of optimized pH gradient systems for the separation of charge variants of trastuzumab after forced degradation study. pH gradient-based elution resulted in high-resolution separation of some 20 charge variants after 3 weeks at 37°C under physiological conditions. The charge variants were further characterized by LC-MS-based peptide mapping. There was no significant difference in the binding properties to HER2 or a range of Fcγ receptors between non-stressed and stressed trastuzumab.

AB - Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and in vivo because of the susceptibility of monoclonal antibodies to undergo chemical modifications. Modifications may adversely affect the potency of the drug, induce immunogenicity or affect pharmacokinetics. In this study, we evaluated the application of optimized pH gradient systems for the separation of charge variants of trastuzumab after forced degradation study. pH gradient-based elution resulted in high-resolution separation of some 20 charge variants after 3 weeks at 37°C under physiological conditions. The charge variants were further characterized by LC-MS-based peptide mapping. There was no significant difference in the binding properties to HER2 or a range of Fcγ receptors between non-stressed and stressed trastuzumab.

UR - http://europepmc.org/abstract/med/34492576

U2 - 10.1016/j.chroma.2021.462506

DO - 10.1016/j.chroma.2021.462506

M3 - Article

C2 - 34492576

SN - 1873-3778

VL - 1655

JO - Journal of chromatography A

JF - Journal of chromatography A

M1 - 462506

ER -

Change of charge variant composition of trastuzumab upon stressing at physiological conditions

Abstract

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