Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

Radhakrishnan, Devesh; Robinson, Anne S.; Ogunnaike, Babatunde A.

doi:10.3390/antib7010001

Cited by 26 publications

(23 citation statements)

References 40 publications

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“…Modeling efforts have furthermore allowed for the prediction of glycosylation patterns under these same feeding treatments (Villiger, Scibona et al, ). A separate study similarly concluded that timewise supplementation of manganese is critical for glycosylation control in mAbs (Radhakrishnan, Robinson & Ogunnaike, ). During both lag and exponential phases, additional manganese supplementation demonstrated a more significant impact on fucosylated glycans than during the stationary phase.…”

Section: Impact Of Trace Metal Supplementation On Product Qualitymentioning

confidence: 94%

“…This examination concurrently focused on metal chelation using ethylenediaminetetraacetic acid (EDTA) in a design of experiments to further demonstrate the extent of controllable glycosylation. When supplied in the presence of additional manganese, EDTA supplementation showed to enhance both cell growth and mAb titer alike, as well as alter the distribution of glycans (Radhakrishnan et al, ). Because EDTA can chelate any number of trace metals in cell culture media, a unique relationship between excess manganese and adjusted trace metal availability by EDTA is presented.…”

Section: Impact Of Trace Metal Supplementation On Product Qualitymentioning

confidence: 99%

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Consequences of trace metal variability and supplementation on Chinese hamster ovary (CHO) cell culture performance: A review of key mechanisms and considerations

Graham

Bhatia

Yoon

2019

Biotech & Bioengineering

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Trace metals are supplied to chemically-defined media (CDM) for optimal Chinese hamster ovary (CHO) cell culture performance during the production of monoclonal antibodies and other therapeutic proteins. However, lot-to-lot and vendor-to-vendor variability in raw materials consequently leads to an imbalance of trace metals that are supplied to CDM. This imbalance can yield detrimental effects rooted in several primary mechanisms and pathways including oxidative stress, apoptosis, lactate accumulation, and unfavorable glycan synthesis. Recent research endeavors involve supplying zinc, copper, and manganese to CDM in excess to further maximize culture productivity and product quality. These treatments significantly impact critical quality attributes and furthermore highlight the degree to which trace metal availability can affect CHO cell culture performance. This review highlights the role of trace metal variability, supplementation, and interplay on key cellular mechanisms responsible for overall culture performance and the production and quality of therapeutic proteins. K E Y W O R D SCHO cell culture performance, raw material variability, therapeutic proteins, trace metal deficiency, trace metal supplementation, trace metals

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Section: Impact Of Trace Metal Supplementation On Product Qualitymentioning

confidence: 94%

Section: Impact Of Trace Metal Supplementation On Product Qualitymentioning

confidence: 99%

Consequences of trace metal variability and supplementation on Chinese hamster ovary (CHO) cell culture performance: A review of key mechanisms and considerations

Graham

Bhatia

Yoon

2019

Biotech & Bioengineering

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“…Because of the importance of the N-linked glycosylation, the majority of recombinant mAb drugs are produced using mammalian host cells in order to achieve human-like glycan structures [22]. In particular, Chinese Hamster Ovary (CHO) cells have become the major expression host for the biopharmaceutical production of therapeutic mAbs [23].The N-linked glycosylation of mAbs has been shown to depend on the host genetic background [24], expression vector [25], media composition [16], media supplements [26], and bioprocess parameters [27]. However, the mechanism of the above dependence remains to be established [28].…”

mentioning

confidence: 99%

“…The N-linked glycosylation of mAbs has been shown to depend on the host genetic background [24], expression vector [25], media composition [16], media supplements [26], and bioprocess parameters [27]. However, the mechanism of the above dependence remains to be established [28].…”

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confidence: 99%

Glycosylation Flux Analysis of Immunoglobulin G in Chinese Hamster Ovary Perfusion Cell Culture

et al. 2018

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The terminal sugar molecules of the N-linked glycan attached to the fragment crystalizable (Fc) region is a critical quality attribute of therapeutic monoclonal antibodies (mAbs) such as immunoglobulin G (IgG). There exists naturally-occurring heterogeneity in the N-linked glycan structure of mAbs, and such heterogeneity has a significant influence on the clinical safety and efficacy of mAb drugs. We previously proposed a constraint-based modeling method called glycosylation flux analysis (GFA) to characterize the rates (fluxes) of intracellular glycosylation reactions. One contribution of this work is a significant improvement in the computational efficiency of the GFA, which is beneficial for analyzing large datasets. Another contribution of our study is the analysis of IgG glycosylation in continuous perfusion Chinese Hamster Ovary (CHO) cell cultures. The GFA of the perfusion cell culture data indicated that the dynamical changes of IgG glycan heterogeneity are mostly attributed to alterations in the galactosylation flux activity. By using a random forest regression analysis of the IgG galactosylation flux activity, we were further able to link the dynamics of galactosylation with two process parameters: cell-specific productivity of IgG and extracellular ammonia concentration. The characteristics of IgG galactosylation dynamics agree well with what we previously reported for fed-batch cultivations of the same CHO cell strain.Processes 2018, 6, 176 2 of 15 and process analytical technology put further emphasis on implementing quantitative approaches for process improvements in biopharmaceutical manufacturing [4,5].Continuous manufacturing technology offers an effective and flexible way for the large scale and robust production of drug compounds [6]. In the biopharmaceutical industry, the application of continuous cell culture technology has thus far been limited to the production of unstable products that require constant recovery [7]. Nevertheless, continuous perfusion cell cultures have previously been demonstrated to be capable of producing antibodies at a volumetric rate that matches or exceeds that of fed-batch cultures [8]. In addition to the high productivity, the stable steady state operation mode in conjunction with the short residence time of perfusion cultures translates to a tight maintenance of product quality. Whether or not product qualities and their key controlling parameters can be directly translated from the traditional batch/fed-batch cell cultures to the perfusion cell cultivations is still unresolved. Past work on converting mAb production from batch/fed-batch to perfusion cell cultures gave conflicting reports on product qualities, where a few studies demonstrated consistent product qualities [9,10], and many others showed differences between the two production modes [11][12][13][14][15].Among the key critical quality attributes (CQAs) of therapeutic mAbs is the glycan structures of the Fc domain [16]. The N-linked glycosylation is a common post-translational modification of proteins, ...

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“…Finally, Zobel-Roos et al [9] propose a process analytical approach allowing for controlled automation of the downstream process by inline concentration measurements based on UV/VIS spectral analysis. In the same area, Radhakrishnan and colleagues show how time-dependent media supplementation by MnCl2 can be used to control the glycosylation profile of antibodies [10]. Castellanos and colleagues use small-angle scattering (SAS) combined with size-exclusion multi-angle light scattering high-performance liquid chromatography and molecular modeling to characterize antibody-antigen complexes in solution [11].…”

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confidence: 99%

Special Issue: Monoclonal Antibodies

Klein

2018

Antibodies

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Monoclonal antibodies are utilized in clinical practice for the treatment of various diseases including cancer, autoimmunity, metabolic and infectious diseases. Over the last 20 years monoclonal antibodies have established themselves as therapeutics and various so-called "blockbuster" drugs are in fact antibodies. Currently ca. 80 monoclonal antibodies have been approved in Europe and the US and several hundreds of them are currently in early and advanced clinical trials. Notably, the field has significantly advanced in the last decade, and nowadays, a large proportion of antibodies in development are engineered antibodies including bispecific antibodies, antibody drug conjugates and novel antibody-like scaffolds. This Special Issue on "Monoclonal Antibodies" includes original manuscripts and reviews covering various aspects related to the discovery, analytical characterization, manufacturing and development of therapeutic and engineered antibodies.The collection starts with a number of reviews. Cho and colleagues [1] review the state of the art in the therapy of multiple myeloma where antibody-based immunotherapies are changing the current treatment paradigm, and Wang-Lin and Balthasar summarize pharmacokinetic and pharmacodynamic considerations that are important for the treatment of bacterial infections by monoclonal antibodies [2]. Finally, Fülöp and colleagues review the role of complement activation in infusion reactions associated with the application of monoclonal antibodies and the potential use of complement factor H for its prevention [3].A series of original articles describes novel monoclonal antibodies for potential diagnostic or therapeutic application. Rashidian and colleagues describe a novel rabbit monoclonal antibody MRQ-67 that specifically recognize the R132H mutation of isocitrate dehydrogenase 1 (IDH1) which are prevalent in diffuse astrocytomas, oligodendrogliomas, and secondary glioblastomas but not the wildtype IDH1. MRQ-67 is able to identify neoplastic cells in glioma tissue specimens and can be used as a tool in glioma subtyping [4]. Zhang and colleagues have identified novel monoclonal antibodies against the Plasmodium falciparum circumsporozoite protein that is a major and immunodominant protective antigen on the surface of plasmodium sporozoites [5]. These antibodies are specific for the central repeat region and mediate protection against challenges from sporozoites. Finally, Rocha and colleagues generated antibodies directed against novel epitopes of the Dengue nonstructural protein 1 (NS1) which is a multi-functional glycoprotein essential for viral replication and modulation of host innate immune responses and represents a surrogate marker for infection [6]. These antibodies are able to differentiate Dengue and Zika virus infections and may contribute to the development of novel diagnostic tools.In a series of three articles, Strube and colleagues [7][8][9] describe approaches useful for the manufacturing and analytical characterization of monoclonal antibodies. An article by ...

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Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

Cited by 26 publications

References 40 publications

Consequences of trace metal variability and supplementation on Chinese hamster ovary (CHO) cell culture performance: A review of key mechanisms and considerations

Consequences of trace metal variability and supplementation on Chinese hamster ovary (CHO) cell culture performance: A review of key mechanisms and considerations

Glycosylation Flux Analysis of Immunoglobulin G in Chinese Hamster Ovary Perfusion Cell Culture

Special Issue: Monoclonal Antibodies

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