Effects and mechanism of small molecule additives on recombinant protein in CHO cells

Zhang, Qiuli; Mi, Chunliu; Wang, Tianyun

doi:10.1007/s00253-023-12486-4

Cited by 7 publications

(3 citation statements)

References 67 publications

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“…Developing and optimizing cell culture media is a tedious and time-consuming process, compared to normal biochemical reactions due to the complex interaction of cells with media components. Media components are known to modulate critical quality attributes (CQAs) like glycosylation and charge variants (Xie et al 2016 ; Rathore et al 2017 ; Miao et al 2017 ; Gangwar et al 2021 , 2022 ; Romanova et al 2022 ; Zhang et al 2023 ). In this study, we have demonstrated the application of an ML-based approach involving feature selection, prediction, and optimization to identify metal ions and their concentrations for achieving charge variant composition of the innovator product.…”

Section: Discussionmentioning

confidence: 99%

Explainable AI for CHO cell culture media optimization and prediction of critical quality attribute

Gangwar,

Balraj,

Rathore

2024

Appl Microbiol Biotechnol

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Cell culture media play a critical role in cell growth and propagation by providing a substrate; media components can also modulate the critical quality attributes (CQAs). However, the inherent complexity of the cell culture media makes unraveling the impact of the various media components on cell growth and CQAs non-trivial. In this study, we demonstrate an end-to-end machine learning framework for media component selection and prediction of CQAs. The preliminary dataset for feature selection was generated by performing CHO-GS (-/-) cell culture in media formulations with varying metal ion concentrations. Acidic and basic charge variant composition of the innovator product (24.97 ± 0.54% acidic and 11.41 ± 1.44% basic) was chosen as the target variable to evaluate the media formulations. Pearson’s correlation coefficient and random forest-based techniques were used for feature ranking and feature selection for the prediction of acidic and basic charge variants. Furthermore, a global interpretation analysis using SHapley Additive exPlanations was utilized to select optimal features by evaluating the contributions of each feature in the extracted vectors. Finally, the medium combinations were predicted by employing fifteen different regression models and utilizing a grid search and random search cross-validation for hyperparameter optimization. Experimental results demonstrate that Fe and Zn significantly impact the charge variant profile. This study aims to offer insights that are pertinent to both innovators seeking to establish a complete pipeline for media development and optimization and biosimilar-based manufacturers who strive to demonstrate the analytical and functional biosimilarity of their products to the innovator. Key points • Developed a framework for optimizing media components and prediction of CQA. • SHAP enhances global interpretability, aiding informed decision-making. • Fifteen regression models were employed to predict medium combinations.

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Section: Discussionmentioning

confidence: 99%

Explainable AI for CHO cell culture media optimization and prediction of critical quality attribute

Gangwar,

Balraj,

Rathore

2024

Appl Microbiol Biotechnol

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“…Another way to control protease activity in cell culture is to add inhibitors or alternative substrates (Table 4 ), such as plant peptone or albumin to the medium (Li et al 2021 ; Schauer et al 2018 ; Zhang et al 2023b ). With developing quality by design (QbD) approach, the researchers revealed many physical and chemical factors that affect the mass of POIs, and screened for the unique substances at one or more stages of the RTP development process, to weaken degradation through combining with the properties of the target protein.…”

Section: Overcome Strategies To Rtp Degradationmentioning

confidence: 99%

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

Geng,

Zhao,

Zhang

et al. 2024

Appl Microbiol Biotechnol

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Mammalian cell lines are frequently used as the preferred host cells for producing recombinant therapeutic proteins (RTPs) having post-translational modified modification similar to those observed in proteins produced by human cells. Nowadays, most RTPs approved for marketing are produced in Chinese hamster ovary (CHO) cells. Recombinant therapeutic antibodies are among the most important and promising RTPs for biomedical applications. One of the issues that occurs during development of RTPs is their degradation, which caused by a variety of factors and reducing quality of RTPs. RTP degradation is especially concerning as they could result in reduced biological functions (antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity) and generate potentially immunogenic species. Therefore, the mechanisms underlying RTP degradation and strategies for avoiding degradation have regained an interest from academia and industry. In this review, we outline recent progress in this field, with a focus on factors that cause degradation during RTP production and the development of strategies for overcoming RTP degradation. Key points • The recombinant therapeutic protein degradation in CHO cell systems is reviewed. • Enzymatic factors and non-enzymatic methods influence recombinant therapeutic protein degradation. • Reducing the degradation can improve the quality of recombinant therapeutic proteins.

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“…The mechanisms by which serum alternatives and other chemical supplements enhance the production of recombinant proteins are multifaceted, encompassing the entire process of exogenous genes from transcription and translation to protein folding and assembly to secretion. [ 107 ] Current researches have revealed that animal‐free extracts can significantly increase the synthesis efficiency of the recombinant proteins by increasing cell viability as well as modulating relevant signaling pathways. Some small molecule organic acids are able to activate transcription factors through epigenetic modifications and thus affect gene expression and regulation.…”

Section: Mechanisms Of Sfm On Cell Growth and Recombinant Protein Pro...mentioning

confidence: 99%

Advances in serum‐free media for CHO cells: From traditional serum substitutes to microbial‐derived substances

Zhang,

Zhao,

et al. 2024

Biotechnology Journal

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The Chinese hamster ovary (CHO) cell is an epithelial‐like cell that produces proteins with post‐translational modifications similar to human glycosylation. It is widely used in the production of recombinant therapeutic proteins and monoclonal antibodies. Culturing CHO cells typically requires the addition of a certain proportion of fetal bovine serum (FBS) to maintain cell proliferation and passaging. However, serum is characterized by its complex composition, batch‐to‐batch variability, high cost, and potential risk of exogenous contaminants such as mycoplasma and viruses, which impact the purity and safety of the synthesized proteins. Therefore, search for serum alternatives and development of serum‐free media for CHO‐based protein biomanufacturing are of great significance. This review systematically summarizes the application advantages of CHO cells and strategies for high‐density expression. It highlights the developmental trends of serum substitutes from human platelet lysates to animal‐free extracts and microbial‐derived substances and elucidates the mechanisms by which these substitutes enhance CHO cell culture performance and recombinant protein production, aiming to provide theoretical guidance for exploring novel serum alternatives and developing serum‐free media for CHO cells.

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Effects and mechanism of small molecule additives on recombinant protein in CHO cells

Cited by 7 publications

References 67 publications

Explainable AI for CHO cell culture media optimization and prediction of critical quality attribute

Explainable AI for CHO cell culture media optimization and prediction of critical quality attribute

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

Advances in serum‐free media for CHO cells: From traditional serum substitutes to microbial‐derived substances

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