Small-molecule control of antibody N-glycosylation in engineered mammalian cells

Chang, Michelle M.; Gaidukov, Leonid; Jung, Gueyoung; Tseng, Wen Allen; Scarcelli, John J.; Cornell, Richard J.; Marshall, Jeffrey; Lyles, Jonathan L.; Sakorafas, Paul; Chu, An-Hsiang Adam; Cote, Kaffa; Tzvetkova, Boriana; Dolatshahi, Sepideh; Sumit, Madhuresh; Mulukutla, Bhanu Chandra; Lauffenburger, Douglas A.; Figueroa, Bruno; Summers, Nevin M.; Lu, Timothy K.; Weiss, Ron

doi:10.1038/s41589-019-0288-4

Cited by 61 publications

(41 citation statements)

References 51 publications

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“…Additionally, a multi-omics study was carried out on the impact of cysteine feed level on cell viability and IgG 1 mAb production in 5 l bioreactors using CHO cells so as to obtain an in-depth understanding of the CHO cell biology (Ali et al, 2019). In a recent study, CHO cells were engineered with synthetic genetic circuits to tune the N-glycosylation of a stably expressed IgG (Chang et al, 2019).…”

Section: Mammalian Cellsmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

397

243

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Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

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Section: Mammalian Cellsmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

397

243

View full text Add to dashboard Cite

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“…1 B, right) [ 28 , 31 ]. Very recently, Weiss’s group [ 32 ] engineered Chinese hamster ovary (CHO) cells with synthetic genetic circuits to rationally tune N -glycosylation of a stably expressed IgG under small-molecule inducible promoters. With the help of a bioorthogonal small-molecule inducer, they achieved stable expression of the glycosyltransferase genes at tunable levels, allowing for a wide range of galactosylated and fucosylated species not easily accessible before.…”

Section: Analysis Of Molecular Structurementioning

confidence: 99%

Recent progress in the molecular imaging of therapeutic monoclonal antibodies

Zeng

Qian

2020

Journal of Pharmaceutical Analysis

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Therapeutic monoclonal antibodies have become one of the central components of the healthcare system and continuous efforts are made to bring innovative antibody therapeutics to patients in need. It is equally critical to acquire sufficient knowledge of their molecular structure and biological functions to ensure the efficacy and safety by incorporating new detection approaches since new challenges like individual differences and resistance are presented. Conventional techniques for determining antibody disposition including plasma drug concentration measurements using LC-MS or ELISA, and tissue distribution using immunohistochemistry and immunofluorescence are now complemented with molecular imaging modalities like positron emission tomography and near-infrared fluorescence imaging to obtain more dynamic information, while methods for characterization of antibody’s interaction with the target antigen as well as visualization of its cellular and intercellular behavior are still under development. Recent progress in detecting therapeutic antibodies, in particular, the development of methods suitable for illustrating the molecular dynamics, is described here.

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“…The term “synthetic glycobiology” was first used to describe the redesign of GT assembly lines for the production of specific glycan structures using protein engineering and chemical approaches (Czlapinski and Bertozzi, 2006 ). This initial definition referred narrowly to the exploitation of Golgi-resident GTs to engineer protein glycosylation inside and on the surface of eukaryotic cells, as exemplified by a number of notable glycoengineering studies in yeast (Choi et al, 2003 ; Hamilton et al, 2003 ) and more recently in mammalian cells (Meuris et al, 2014 ; Chang et al, 2019 ). These successes notwithstanding, simpler, cell-viability independent systems that permit bottom-up assembly of prescribed glycosylation pathways and controllable biosynthesis of designer glycomolecules are of great scientific and technological interest, and have the potential to be transformative.…”

Section: Synthetic Glycobiologymentioning

confidence: 99%

“…Such knowledge, in turn, provided a blueprint for genetic reconstruction of CHO cells with desirable glycosylation capacities including those producing human-like α2,6-linked sialic acid-capped glycoforms on therapeutic proteins such as human IgG and Erythropoietin (EPO) (Yang et al, 2015 ; Caval et al, 2018 ; Schulz et al, 2018 ). Another notable example from the Weiss group explored the use of CRISPR/Cas9 to implement synthetic gene circuits in CHO cells, allowing tunable N -glycan profiles of CHO culture-derived IgGs in a small molecule concentration-dependent manner (Chang et al, 2019 ). More recently, precision gene editing was used to create a library of validated CRISPR/Cas9 guide RNA targeting constructs for all human GT genes (Narimatsu et al, 2018 ).…”

Section: Cell-based and Cell-free Biosynthesis Of Structurally-definementioning

confidence: 99%

Cell-Free Synthetic Glycobiology: Designing and Engineering Glycomolecules Outside of Living Cells

Jaroentomeechai

Taw

et al. 2020

Front. Chem.

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Glycans and glycosylated biomolecules are directly involved in almost every biological process as well as the etiology of most major diseases. Hence, glycoscience knowledge is essential to efforts aimed at addressing fundamental challenges in understanding and improving human health, protecting the environment and enhancing energy security, and developing renewable and sustainable resources that can serve as the source of next-generation materials. While much progress has been made, there remains an urgent need for new tools that can overexpress structurally uniform glycans and glycoconjugates in the quantities needed for characterization and that can be used to mechanistically dissect the enzymatic reactions and multi-enzyme assembly lines that promote their construction. To address this technology gap, cell-free synthetic glycobiology has emerged as a simplified and highly modular framework to investigate, prototype, and engineer pathways for glycan biosynthesis and biomolecule glycosylation outside the confines of living cells. From nucleotide sugars to complex glycoproteins, we summarize here recent efforts that harness the power of cell-free approaches to design, build, test, and utilize glyco-enzyme reaction networks that produce desired glycomolecules in a predictable and controllable manner. We also highlight novel cell-free methods for shedding light on poorly understood aspects of diverse glycosylation processes and engineering these processes toward desired outcomes. Taken together, cell-free synthetic glycobiology represents a promising set of tools and techniques for accelerating basic glycoscience research (e.g., deciphering the “glycan code”) and its application (e.g., biomanufacturing high-value glycomolecules on demand).

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Small-molecule control of antibody N-glycosylation in engineered mammalian cells

Cited by 61 publications

References 51 publications

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Recent progress in the molecular imaging of therapeutic monoclonal antibodies

Cell-Free Synthetic Glycobiology: Designing and Engineering Glycomolecules Outside of Living Cells

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