Charge Variants Analysis of Recombinant Monoclonal Antibodies

Torkashvand, Fatemeh; Vaziri, Behrouz

doi:10.26717/bjstr.2019.20.003395

Cited by 3 publications

(1 citation statement)

References 17 publications

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“…Since mAbs charge variants are not similar in their impacts on safety and efficacy [ 11 ] , determining these effects is vital to define the design space in the downstream process of each mAb. In this regard, it seems that the charge variant profile of each therapeutic mAb is unique and it is a key factor for the consistency in the production process [ 27 ] .…”

Section: Discussionmentioning

confidence: 99%

Trastuzumab Charge Variants: a Study on Physicochemical and Pharmacokinetic Properties

Torkashvand,

Mehranfar,

Rashidi Gero

et al. 2023

IBJ

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Background: Post-translational modifications in bioprocessing and storage of recombinant mAbs are the main sources of charge variants. While the profile of these kinds of variants is considered an important quality attribute of therapeutic mAbs, there is controversy about their direct role on safety and efficacy. In this study, the physicochemical and PK properties of separated charge variants belonging to a trastuzumab potential biosimilar were examined. Methods: The APs, BPs, and main variants of trastuzumab were separated and enriched by semi-preparative WCX. A panel of analytical techniques was utilized to characterize the physicochemical properties of these variants. The binding affinity to HER2 and FcγRs and the PK parameters were evaluated for each variant. Results: Based on the results, the charge variants of the proposed biosimilar had no significant influence on the examined efficacy and PK parameters. Conclusion: During the development and production of biosimilar mAbs, studying the effects of their charge variants on efficacy and PK parameters is needed.

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Section: Discussionmentioning

confidence: 99%

Trastuzumab Charge Variants: a Study on Physicochemical and Pharmacokinetic Properties

Torkashvand,

Mehranfar,

Rashidi Gero

et al. 2023

IBJ

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Structural and Functional Analysis of CEX Fractions Collected from a Novel Avastin® Biosimilar Candidate and Its Innovator: A Comparative Study

Gürel

Berksoz²,

Çapkın

et al. 2022

Pharmaceutics

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Avastin® is a humanized recombinant monoclonal antibody used to treat cancer by targeting VEGF-A to inhibit angiogenesis. SIMAB054, an Avastin® biosimilar candidate developed in this study, showed a different charge variant profile than its innovator. Thus, it is fractionated into acidic, main, and basic isoforms and collected physically by Cation Exchange Chromatography (CEX) for a comprehensive structural and functional analysis. The innovator product, fractionated into the same species and collected by the same method, is used as a reference for comparative analysis. Ultra-Performance Liquid Chromatography (UPLC) ESI-QToF was used to analyze the modifications leading to charge heterogeneities at intact protein and peptide levels. The C-terminal lysine clipping and glycosylation profiles of the samples were monitored by intact mAb analysis. The post-translational modifications, including oxidation, deamidation, and N-terminal pyroglutamic acid formation, were determined by peptide mapping analysis in the selected signal peptides. The relative binding affinities of the fractionated charge isoforms against the antigen, VEGF-A, and the neonatal receptor, FcRn, were revealed by Surface Plasmon Resonance (SPR) studies. The results show that all CEX fractions from the innovator product and the SIMAB054 shared the same structural variants, albeit in different ratios. Common glycoforms and post-translational modifications were the same, but at different percentages for some samples. The dissimilarities were mostly originating from the presence of extra C-term Lysin residues, which are prone to enzymatic degradation in the body, and thus they were previously assessed as clinically irrelevant. Another critical finding was the presence of different glyco proteoforms in different charge species, such as increased galactosylation in the acidic and afucosylation in the basic species. SPR characterization of the isolated charge variants further confirmed that basic species found in the CEX analyses of the biosimilar candidate were also present in the innovator product, although at lower amounts. The charge variants’ in vitro antigen- and neonatal receptor-binding activities varied amongst the samples, which could be further investigated in vivo with a larger sample set to reveal the impact on the pharmacokinetics of drug candidates. Minor structural differences may explain antigen-binding differences in the isolated charge variants, which is a key parameter in a comparability exercise. Consequently, such a biosimilar candidate may not comply with high regulatory standards unless the binding differences observed are justified and demonstrated not to have any clinical impact.

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Trastuzumab Charge Variants: a Study on Physicochemical and Pharmacokinetic Properties

Torkashvand,

Mehranfar,

Rashidi Gero

et al. 2023

ibj

Self Cite

View full text Add to dashboard Cite

Background: Post-translational modifications in bioprocessing and storage of recombinant mAbs are the main sources of charge variants. While the profile of these kinds of variants is considered an important quality attribute of therapeutic mAbs, there is controversy about their direct role on safety and efficacy. In this study, the physicochemical and PK properties of separated charge variants belonging to a trastuzumab potential biosimilar were examined. Methods: The APs, BPs, and main variants of trastuzumab were separated and enriched by semi-preparative WCX. A panel of analytical techniques was utilized to characterize the physicochemical properties of these variants. The binding affinity to HER2 and FcγRs and the PK parameters were evaluated for each variant. Results: Based on the results, the charge variants of the proposed biosimilar had no significant influence on the examined efficacy and PK parameters. Conclusion:During the development and production of biosimilar mAbs, studying the effects of their charge variants on efficacy and PK parameters is needed.

show abstract

Charge Variants Analysis of Recombinant Monoclonal Antibodies

Cited by 3 publications

References 17 publications

Trastuzumab Charge Variants: a Study on Physicochemical and Pharmacokinetic Properties

Trastuzumab Charge Variants: a Study on Physicochemical and Pharmacokinetic Properties

Structural and Functional Analysis of CEX Fractions Collected from a Novel Avastin® Biosimilar Candidate and Its Innovator: A Comparative Study

Trastuzumab Charge Variants: a Study on Physicochemical and Pharmacokinetic Properties

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