Flux Sampling Suggests Metabolic Signatures of High Antibody‐Producing CHO Cells

Biotechnology and Bioengineering, Journal Year: 2025, Volume and Issue: unknown

Published: April 11, 2025

Chinese hamster ovary (CHO) cells remain the industry standard for producing numerous therapeutic proteins, particularly monoclonal antibodies (mAbs). However, achieving higher recombinant protein titers remains an ongoing challenge and a fundamental understanding of cellular mechanism driving improved bioprocess performance elusive. To directly address these challenges achieve substantial improvements, more in-depth function within environment may be required. Over past decade, significant advancements have been made in building genome-scale metabolic models (GEMs) CHO cells, bridging gap between high information content 'omics data ability to perform silico phenotypic predictions. Here, time-course transcriptomics has employed constrain culture phase-specific GEMs, representing early exponential, late stationary/death phases cell fed-batch bioreactor culture. Temporal data, including metabolite uptake secretion rates, as well growth productivity, used validate flux sampling results. Additionally, mAb-producing solutions identified signatures associated with mAb production hypothesized. Finally, constraint-based modeling utilized infer specific amino acids, cysteine, histidine, leucine, isoleucine, asparagine, serine, which could drive increased guide optimal media feed formulations.

Language: Английский

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m6A modification profiles of the CHO cells with differential recombinant protein expression using MeRIP-seq/RNA-seq

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: unknown, P. 143429 - 143429

Published: April 1, 2025

Language: Английский

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Modeling and Optimization of Recombinant Tocilizumab Production From Pichia pastoris Using Response Surface Methodology and Artificial Neural Network

Biotechnology and Bioengineering, Journal Year: 2025, Volume and Issue: unknown

Published: May 15, 2025

ABSTRACT This study has demonstrated the optimization of defined medium that significantly enhanced production recombinant monoclonal antibody (mAb) Tocilizumab (TCZ) as full‐length and Fab fragment from Pichia pastoris . Out four tested media, FM22 was found to be suitable for growth strains yield. Among various carbon nitrogen sources tested, mannitol glycine, respectively, were TCZ. Similarly, sorbitol ammonium sulfate sources, Fab. The components influenced TCZ mannitol, histidine, K 2 SO 4 sorbitol, sulfate, KH PO , CaSO .2H O Fab, using a two‐level factorial Plackett‐Burman design. screened optimized response surface methodology (Box‐Behnken Design). Artificial neural network (ANN) models combined with genetic algorithms (GA) further improved predictions showed remarkable impact on mAb in P. Under optimal levels components, determined 0.35 mg/L 0.42 g/L, shake‐flask culture. yield batch reactor (2‐L culture) 0.44 0.45 at components. overall increased yields observed 3.8 2.9‐folds respectively.

Language: Английский

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Characterization and design of dipeptide media formulation for scalable therapeutic production

Applied Microbiology and Biotechnology, Journal Year: 2025, Volume and Issue: 109(1)

Published: Jan. 14, 2025

Process intensification and simplification in biopharmaceutical manufacturing have driven the exploration of advanced feeding strategies to improve culture performance process consistency. Conventional media design strategies, however, are often constrained by stability solubility challenges amino acids, particularly large-scale applications. As a result, dipeptides emerged as promising alternatives. Despite extensive research on dipeptide supplementation Chinese hamster ovary (CHO) cell-based has received comparatively less attention. In this review, we critically analyze associated with acids prone instability poor (e.g., glutamine, cysteine, tyrosine), explore potential address these limitations. We intricate mechanisms transport enzymatic cleavage, highlighting how chemical properties, stereoisomerism, competitive metabolites influence their utilization. Notably, while most exhibit enhanced solubility, stabilization effects remain variable, underlining need for rational design. To guide future innovations, propose tailored derived specific biomanufacturing needs integrating multi-omics analysis, metabolic flux modeling, artificial intelligence (AI) modeling. KEY POINTS : •Explored solution acid enhancing cell performance. •Discussed transporter kinetics cleavage enzymes influencing utilization biomanufacturing. •Suggested various identifying appropriate pairs bioprocess efficiency.

Language: Английский

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Single-B-cell cloning and recombinant antibodies generation to analyze the antigenicity of porcine reproductive and respiratory syndrome virus nonstructural protein 12

Veterinary Microbiology, Journal Year: 2025, Volume and Issue: 304, P. 110506 - 110506

Published: April 8, 2025

Language: Английский

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