Evaluation of Heavy-Chain C-Terminal Deletion on Product Quality and Pharmacokinetics of Monoclonal Antibodies

Jiang, Guoying; Yu, Christopher; Yadav, Daniela Bumbaca; Hu, Zhilan; Amurao, Annamarie; Duenas, Eileen; Wong, Margaret Wan Nar; Iverson, Mark; Zheng, Kai; Lam, Xanthe M.; Chen, Jia; Vega, Roxanne; Ulufatu, Sheila; Leddy, Cecilia; Davis, Helen; Shen, Amy; Wong, Pin Yee; Harris, Reed J.; Wang, Y. John; Li, Dongwei

doi:10.1016/j.xphs.2016.04.027

Cited by 27 publications

(30 citation statements)

References 19 publications

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“…Protein products with pI difference greater than one will present measurable changes in tissue distribution and kinetics (Boswell et al, ). It is also important to note that while highly purified and modified forms of charge variants display substantial pharmacokinetic (PK) changes, the effect on the overall mAb pool is typically limited and insignificant at reported literature values, ∼25 ± 5% acidic variants (Jiang et al, ; Khawli et al, ; Zhao et al, ).…”

Section: Commonly Occurring Charge Variants and Their Cqa Managementmentioning

confidence: 99%

“…Cell line development has also been particularly effective at controlling C‐terminal Lys. Investigations into the carboxypeptidase activity, the enzyme that cleaves the C‐terminal Lys, allows for targeted modulation by cell line engineering (Dick, Qiu, Mahon, Adamo, & Cheng, ; Hu et al, ), and deletion of C‐terminal amino residues has also successfully controlled the basic charge variants (Hu et al, ; Jiang et al, ). While cell line development may effectively control protein modifications caused by endogenous mechanisms, many charge variant modifications occur due to extracellular interactions in a molecule‐dependent manner that may be difficult to predict and control.…”

Section: Upstream Process Strategies For Charge Variants Controlmentioning

confidence: 99%

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Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Chung

Tian

Tan

et al. 2018

Biotech & Bioengineering

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Controlling the charge profile of therapeutic protein is a critical challenge in the current quality-by-design (QbD) paradigm, throughout all phases of biologics process development (PD): cell line development, upstream cell culture, recovery process, downstream purification, and analytical characterization. Charge variant profiles may influence efficacy and/or lead to unintended side-effects. Thus, maintaining a consistent charge profile is of tremendous importance, and increasingly, researchers have focused efforts toward developing strategies to mitigate variability during cell culture and to improve separation and detection of charge variants. Current understanding of factors affecting charge variant formation during manufacturing remains inadequate, and sometimes, even substantial commitment of resources may still not fully achieve the desired or consistent profiles. As such, this review attempts to provide a comprehensive resource for the biologics community by summarizing the impact of charge variants and CQA management, analytical methods for charge variant detection, as well as strategies in downstream and upstream PD for controlling charge variant profiles.

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Section: Commonly Occurring Charge Variants and Their Cqa Managementmentioning

confidence: 99%

Section: Upstream Process Strategies For Charge Variants Controlmentioning

confidence: 99%

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Chung

Tian

Tan

et al. 2018

Biotech & Bioengineering

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“…In contrast, aspartate‐glycine motives are prone to succinimide formation that increases the p I of the affected species. Also many other modifications like glutamine/pyroglutamate conversion , sialylation of glycan structures , C‐terminal lysine heterogeneity or oxidation of disulfide bonds can influence the charge profile.…”

Section: Introductionmentioning

confidence: 99%

Optimization of capillary zone electrophoresis for charge heterogeneity testing of biopharmaceuticals using enhanced method development principles

et al. 2017

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CZE is a well‐established technique for charge heterogeneity testing of biopharmaceuticals. It is based on the differences between the ratios of net charge and hydrodynamic radius. In an extensive intercompany study, it was recently shown that CZE is very robust and can be easily implemented in labs that did not perform it before. However, individual characteristics of some examined proteins resulted in suboptimal resolution. Therefore, enhanced method development principles were applied here to investigate possibilities for further method optimization. For this purpose, a high number of different method parameters was evaluated with the aim to improve CZE separation. For the relevant parameters, design of experiments (DoE) models were generated and optimized in several ways for different sets of responses like resolution, peak width and number of peaks. In spite of product specific DoE optimization it was found that the resulting combination of optimized parameters did result in significant improvement of separation for 13 out of 16 different antibodies and other molecule formats. These results clearly demonstrate generic applicability of the optimized CZE method. Adaptation to individual molecular properties may sometimes still be required in order to achieve optimal separation but the set screws discussed in this study [mainly pH, identity of the polymer additive (HPC versus HPMC) and the concentrations of additives like acetonitrile, butanolamine and TETA] are expected to significantly reduce the effort for specific optimization.

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“…If however an effector‐less antibody is desired, an aglycosylated –GK antibody may be engineered . While our initial investigations suggested that deletion of C‐terminal lysine or glycine–lysine residues from HC does not affect antibody glycosylation, potency, PK, and bioavailability relative to the WT antibody, nevertheless, the immunogenicity of the glycine–lysine deletion in antibodies will need to be tested in order to ensure patient safety prior to its utilization for commercial production.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, antibody lacking HC C‐terminal lysine should have the same potency as its WT counterpart. Indeed, our prior evaluations did not reveal any differences in stability, pharmacokinetic (PK) properties, or potency amongst WT antibodies and antibodies lacking either HC C‐terminal lysine (‐K) or both lysine and glycine (‐GK) . In this study, we have investigated the impact of HC C‐terminal amino acids on antibody productivity and product quality using three different monoclonal antibody molecules (mAbs).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of heavy chain C‐terminal deletions on productivity and product quality of monoclonal antibodies in Chinese hamster ovary (CHO) cells

Hu¹,

Tang²,

Misaghi³

et al. 2017

Biotechnology Progress

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Monoclonal antibodies (mAbs) have been well established as potent therapeutic agents and are used to treat many different diseases. During cell culture production, antibody charge variants can be generated by cleavage of heavy chain (HC) C-terminal lysine and proline amidation. Differences in levels of charge variants during manufacturing process changes make it challenging to demonstrate process comparability. In order to reduce heterogeneity and achieve consistent product quality, we generated and expressed antibodies with deletion of either HC C-terminal lysine (-K) or lysine and glycine (-GK). Interestingly, clones that express antibodies lacking HC C-terminal lysine (-K) had considerably lower specific productivities compared to clones that expressed either wild type antibodies (WT) or antibodies lacking HC glycine and lysine (-GK). While no measurable differences in antibody HC and LC mRNA levels, glycosylation and secretion were observed, our analysis suggests that the lower specific productivity of clones expressing antibody lacking HC C-terminal lysine was due to slower antibody HC synthesis and faster antibody degradation. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:786-794, 2017.

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Evaluation of Heavy-Chain C-Terminal Deletion on Product Quality and Pharmacokinetics of Monoclonal Antibodies

Cited by 27 publications

References 19 publications

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Optimization of capillary zone electrophoresis for charge heterogeneity testing of biopharmaceuticals using enhanced method development principles

Evaluation of heavy chain C‐terminal deletions on productivity and product quality of monoclonal antibodies in Chinese hamster ovary (CHO) cells

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