Strategies to enhance productivity and modify product quality in therapeutic proteins

Radhakrishnan, Devesh; Wells, Evan A.; Robinson, Anne S.

doi:10.1016/j.coche.2018.09.005

Cited by 19 publications

(12 citation statements)

References 38 publications

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“…Despite the two markers are often used for describing best cell performance, the focus of critical parameters is shifted toward increased product quality . Thereby, critical quality attributes have to be defined for each molecule of interest individually and quality characteristics are predominantly defined by the process operation mode/conditions and harvest time …”

Section: Resultsmentioning

confidence: 99%

“…[24][25][26] Thereby, critical quality attributes have to be defined for each molecule of interest individually and quality characteristics are predominantly defined by the process operation mode/conditions and harvest time. 22,[27][28][29][30][31][32][33] In semicontinuous small-scale models, N-glycosylation and charge profiles changed distinctly along different culture harvest times, but not by the addition of the CB1 and CB3 feed supplements (Figure 5d, e). The variation of product quality attributes was higher between different harvest times than between different media.…”

Section: Verification Of Performance By Semicontinuous Small-scale mentioning

confidence: 99%

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Rapid development of clone‐specific, high‐performing perfusion media from established feed supplements

Mayrhofer

Reinhart

Castan³

et al. 2019

Biotechnology Progress

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Perfusion cultivation of recombinant CHO cells is of substantial interest to the biopharmaceutical industry. This is due to increased space–time‐yields (STYs) and a short residence time of the recombinant protein in the bioreactor. Economic processes rely on cultivation media supporting rapid growth in the exponential phase and high protein production in the stationary phase at minimal media consumption rates. To develop clone‐specific, high‐performing perfusion media we present a straightforward and rapid two‐step approach combining commercially available basal media and feed supplements using design‐of‐experiment. First, the best performing feed supplements are selected in batch cultures. Then, the mixing ratio of selected feed supplements is optimized in small‐scale semicontinuous perfusion cultures. The final media formulation is supported by statistical response surface modeling of a set of cultivation experiments with blended media formulations. Two best performing novel media blends were finally applied to perfusion bioreactor verification runs to reach 200 × 106 c/ml within 2 weeks at minimum cell‐specific perfusion rates as low as 10–30 pL/c/d. Obtained STYs of 0.4–1.2 g/L/d represent a 10‐fold increase compared to batch cultures. This general workflow is universally applicable to any perfusion platform combining a specific cell line, basal medium, and established feed solutions.

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Section: Resultsmentioning

confidence: 99%

Section: Verification Of Performance By Semicontinuous Small-scale mentioning

confidence: 99%

Rapid development of clone‐specific, high‐performing perfusion media from established feed supplements

Mayrhofer

Reinhart

Castan³

et al. 2019

Biotechnology Progress

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“…Nevertheless, owing to cell engineering, the time for the establishment of productive cell lines of fully humanized mAbs has sharply reduced, limiting it to some months, with increased productivities (up to 100 pg/cell day, representing bioreactor titers of nearly 10 g/L), which is presently crucial for the production of anti-SARS-CoV-2 mAbs [ 264 , 269 – 271 ]. Elevated productive-mAb titres have also been achieved by extensive improvements in the production schemes [ 272 ]. Similarly, improvements in the recovery and purification of mAbs have achieved yields of up to 80% of that produced in bioreactors, and consequently, the manufacturing costs of goods have dropped to 20–100 US$ per gram of the active pharmaceutical ingredient [ 270 , 273 ].…”

Section: Remarks On the Production Of Mabs For Covid-19 Treatmentmentioning

confidence: 99%

“…In addition to the quality assurance, it is necessary to demonstrate batch-by-batch reproducibility in bioprocesses. The production of anti-SARS-CoV-2 mAbs in CHO cells can be seen as a feasible strategy for implementation on an industrial scale in conjunction with high-density cultures [ 272 ], single-use technologies [ 267 ], design/selection techniques for highly productive clones [ 264 , 281 ], and bioprocess optimization [ 266 ]. To achieve this, it is necessary that companies with large biotechnological developments provide insights to the bio-pharmacological industry to combat this global pandemic.…”

Section: Remarks On the Production Of Mabs For Covid-19 Treatmentmentioning

confidence: 99%

Integrative overview of antibodies against SARS-CoV-2 and their possible applications in COVID-19 prophylaxis and treatment

et al. 2021

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SARS-CoV-2 is a novel β-coronavirus that caused the COVID-19 pandemic disease, which spread rapidly, infecting more than 134 million people, and killing almost 2.9 million thus far. Based on the urgent need for therapeutic and prophylactic strategies, the identification and characterization of antibodies has been accelerated, since they have been fundamental in treating other viral diseases. Here, we summarized in an integrative manner the present understanding of the immune response and physiopathology caused by SARS-CoV-2, including the activation of the humoral immune response in SARS-CoV-2 infection and therefore, the synthesis of antibodies. Furthermore, we also discussed about the antibodies that can be generated in COVID-19 convalescent sera and their associated clinical studies, including a detailed characterization of a variety of human antibodies and identification of antibodies from other sources, which have powerful neutralizing capacities. Accordingly, the development of effective treatments to mitigate COVID-19 is expected. Finally, we reviewed the challenges faced in producing potential therapeutic antibodies and nanobodies by cell factories at an industrial level while ensuring their quality, efficacy, and safety.

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“…Because of cost and to protect proprietary advantage, biopharmaceutical companies with both in‐house and outsourced media formulations often do not disclose the final recipes of their commercial media products. However, certain supplements have been identified that affect culture growth rates, mAb titers, and final product quality, and these can be used with varying degrees of success to control desired characteristics in differently formulated media (Graham, Bhatia, & Yoon, 2019; Radhakrishnan, Wells, & Robinson, 2018). For instance, 2‐F‐peracetyl fucose (2FP) supplementation has been shown to lower fucosylation in mAbs by up to 75% by inhibiting fucosyltransferase 8 (FUT8; Ehret, Zimmermann, Eichhorn, & Zimmer, 2019; Mishra, Spearman, Donald, Perreault, & Butler, 2020); galactose addition has raised overall antibody galactosylation (St. Amand, Radhakrishnan, Robinson, & Ogunnaike, 2014); and the presence of uridine has raised titer, integrated viable cell density (below 10 mM concentrations), and galactosylation (Ehret et al, 2019; Grainger & James, 2013; Gramer et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Media supplementation for targeted manipulation of monoclonal antibody galactosylation and fucosylation

Wells

Song

Greer

et al. 2020

Biotech & Bioengineering

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Monoclonal antibodies are critically important biologics as the largest class of molecules used to treat cancers, rheumatoid arthritis, and other chronic diseases. Antibody glycosylation is a critical quality attribute that has ramifications for patient safety and physiological efficacy-one that can be modified by such factors as media formulation and process conditions during production. Using a design-ofexperiments approach, we examined the effect of 2-F-peracetyl fucose (2FP), uridine, and galactose on cell growth and metabolism, titer, and gene expression of key glycosylation-related proteins, and report how the glycoform distribution changed from Days 4 to 7 in a batch process used for IgG1 production from Chinese hamster ovary cells. We observed major glycosylation changes upon supplement addition, where the addition of 2FP decreased antibody fucosylation by up to 48%, galactose addition increased galactosylation by up to 21%, and uridine addition decreased fucosylation and increased galactosylation by 6% and 2%, respectively. Despite having major effects on glycosylation, neither galactose nor 2FP significantly affected cell culture growth, metabolism, or titer. Uridine improved peak cell densities by 23% but also reduced titer by ∼30%. The supplements caused significant changes in gene expression by Day 4 of the cultures where 2FP addition significantly reduced fucosyltransferase 8 and nucleotide sugar transporter gene expression (by ∼2-fold), and uridine addition significantly increased expression of UDP-GlcNAcT (SLC35A3) and B4GALT1-6 genes (by 1.5-3-fold). These gene expression data alongside glycosylation, metabolic, and growth data improve our understanding of the cellular mechanisms affected by media supplementation and suggest approaches for modifying antibody glycosylation in antibody production processes. K E Y W O R D S antibody, Chinese hamster ovary, design of experiments 1 | INTRODUCTION Monoclonal antibodies (mAbs) have been very effective in treating numerous types of diseases and have become a driving force behind the growth of the biotherapeutic industry (Kunert & Reinhart, 2016). More than 86 mAbs and mAb derivatives have been approved by the US Food and Drug Administration (FDA) with ∼90 more currently in clinical trials (Kaplon & Reichert, 2019). In 2019, for example, of the 20 FDA-approved biologics, nine of them are antibodies, antibody fragments, or antibody conjugates (Morrison, 2020).

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Strategies to enhance productivity and modify product quality in therapeutic proteins

Cited by 19 publications

References 38 publications

Rapid development of clone‐specific, high‐performing perfusion media from established feed supplements

Rapid development of clone‐specific, high‐performing perfusion media from established feed supplements

Integrative overview of antibodies against SARS-CoV-2 and their possible applications in COVID-19 prophylaxis and treatment

Media supplementation for targeted manipulation of monoclonal antibody galactosylation and fucosylation

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