Factors Affecting the Expression of Recombinant Protein and Improvement Strategies in Chinese Hamster Ovary Cells

Li, Zhengmei; Fan, Zhenlin; Wang, Xiaoyin; Wang, Tianyun

doi:10.3389/fbioe.2022.880155

Cited by 25 publications

(12 citation statements)

References 115 publications

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“…Recombinant protein expression is a staple of molecular biology and the expression yield determines whether a P.O.I. is suitable for experimental purposes [33, 59]. FPs could thus enhance protein expression yields to characterize the functionality of proteins, whilst serving as an intrinsic fluorescent handle.…”

Section: Discussionmentioning

confidence: 99%

Viral envelope proteins fused to multiple distinct fluorescent reporters to probe receptor binding

Tomris,

van der Woude,

de Paiva Droes Rocha

et al. 2023

Preprint

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Enveloped viruses carry one or multiple proteins with receptor binding functionalities. Functional receptors can either be glycans, proteinaceous or both, recombinant protein approaches are instrumental to gain more insight into these binding properties. Visualizing and measuring receptor binding normally entails antibody detection or direct labelling, whereas direct fluorescent fusions are attractive tools in molecular biology. Here we report a suite of different fluorescent fusions, both N- and/or C- terminal, for influenza A virus hemagglutinins and SARS-CoV-2 spike RBD. The proteins contained a total of three or six fluorescent protein barrels and were applied directly to cells to determine receptor binding properties.

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Section: Discussionmentioning

confidence: 99%

Viral envelope proteins fused to multiple distinct fluorescent reporters to probe receptor binding

Tomris,

van der Woude,

de Paiva Droes Rocha

et al. 2023

Preprint

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show abstract

“…The amount of protein produced in stable secreting cell lines can vary dramatically for different target proteins, and the reasons for this are not well understood and are difficult to predict [ 84 ]. Each step of the process must be reasonably efficient and streamlined to achieve active protein secretion: transcription, translocation of mRNA to the ER surface, an appropriate translation rate, proper folding in the ER, glycosylation, and finally, secretion from the cell.…”

Section: The General Strategy Of Recombinant Protein Expression In Ma...mentioning

confidence: 99%

The Production of Complement Inhibitor Proteins in Mammalian Cell Lines—Light at the End of the Tunnel?

Szvetnik,

Tubak

2024

Biomedicines

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Therapeutic recombinant proteins are powerful tools used for the treatment of many detrimental diseases such as diabetes, cancer, multiple sclerosis, rheumatoid arthritis, hepatitis, and many more. Their importance in disease therapy is growing over small molecule drugs because of their advantages like specificity and reduced side effects. However, the large-scale production of certain recombinant proteins is still challenging despite impressive advancements in biomanufacturing. The complement cascade is considered a rich source of drug targets and natural regulator proteins with great therapeutic potential. However, the versatility of such proteins has been hampered by low production rates. The recent discoveries highlighted here may bring definite improvement in the large-scale recombinant production of complement inhibitor proteins or other difficult-to-express proteins in mammalian cell lines.

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“…All these facts complicate the procedure and increase the production costs [ 196 ]. Thus, improvement of stable cell lines has become a clear need, using strategies involving genetic modification, optimization of expression vector and process engineering [ 197 , 198 ]. With the advancement of CHO cell line development and process optimization, yields of some recombinant proteins (such as monoclonal antibodies) have achieved as high as 5 g/L, or even more than 10 g/L [ 199 , 200 ].…”

Section: Conventional Cell Factories For Recombinant Vaccinesmentioning

confidence: 99%

Recombinant vaccines in 2022: a perspective from the cell factory

et al. 2022

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The last big outbreaks of Ebola fever in Africa, the thousands of avian influenza outbreaks across Europe, Asia, North America and Africa, the emergence of monkeypox virus in Europe and specially the COVID-19 pandemics have globally stressed the need for efficient, cost-effective vaccines against infectious diseases. Ideally, they should be based on transversal technologies of wide applicability. In this context, and pushed by the above-mentioned epidemiological needs, new and highly sophisticated DNA-or RNA-based vaccination strategies have been recently developed and applied at large-scale. Being very promising and effective, they still need to be assessed regarding the level of conferred long-term protection. Despite these fast-developing approaches, subunit vaccines, based on recombinant proteins obtained by conventional genetic engineering, still show a wide spectrum of interesting potentialities and an important margin for further development. In the 80’s, the first vaccination attempts with recombinant vaccines consisted in single structural proteins from viral pathogens, administered as soluble plain versions. In contrast, more complex formulations of recombinant antigens with particular geometries are progressively generated and explored in an attempt to mimic the multifaceted set of stimuli offered to the immune system by replicating pathogens. The diversity of recombinant antimicrobial vaccines and vaccine prototypes is revised here considering the cell factory types, through relevant examples of prototypes under development as well as already approved products.

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Factors Affecting the Expression of Recombinant Protein and Improvement Strategies in Chinese Hamster Ovary Cells

Cited by 25 publications

References 115 publications

Viral envelope proteins fused to multiple distinct fluorescent reporters to probe receptor binding

Viral envelope proteins fused to multiple distinct fluorescent reporters to probe receptor binding

The Production of Complement Inhibitor Proteins in Mammalian Cell Lines—Light at the End of the Tunnel?

Recombinant vaccines in 2022: a perspective from the cell factory

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