[Application Sharing] Key Parameters for Antibody Production: Glucose and Amino Acids

In recent decades, the global annual market for recombinant therapeutic proteins has grown significantly. In particular, monoclonal antibodies (mAbs) have opened up new therapeutic avenues for cancer, inflammatory diseases, infectious diseases, and autoimmune disorders, achieving remarkable success in both regulatory approvals and global sales.

2023-07-12

In recent decades, the global annual market for recombinant therapeutic proteins has grown significantly. In particular, monoclonal antibodies ( mAbs ), it has opened up new therapeutic avenues for cancer, inflammatory diseases, infectious diseases, and autoimmune disorders, and has achieved remarkable success in regulatory approvals and global sales. Chinese hamster ovary ( CHO ) Cells are widely used in the production of recombinant antibodies because they exhibit robust growth and have the potential to generate non-immunogenic antibodies with glycosylation patterns similar to those found in humans. Glycosylation plays a crucial role in therapeutic IgG It plays a critical role in the biological characteristics of a molecule, such as effector function, immunogenicity, stability, and clearance rate. Therefore, controlling glycosylation is essential for meeting regulatory requirements and ensuring quality compliance. The heterogeneity of glycan structures at each glycosylation site can vary depending on the stage of biosynthesis—from less mature forms (e.g., non-glycosylated and high-mannose forms) to more mature types (e.g., galactosylated or sialylated forms).

While glycolysis and glutamine metabolism are... CHO Key metabolic pathways of cells. Through glycolysis, CHO Cells consume glucose as the primary carbon source for energy production and produce lactate as the most common metabolic byproduct. Glutamine catabolism is... CHO The primary pathway by which cells absorb organic nitrogen for biomass synthesis, while simultaneously releasing ammonium ions as the main byproduct. Fed-batch cultivation ( fed-batch ) It is widely used in industry for the production of recombinant antibodies. In fed-batch Medium supplemented with regular, corresponding feedings. / Glucose can provide sufficient nutrients to support cell growth and metabolism, enabling high-density cells to survive and express desired products, thereby extending cell viability and inducing high productivity. However, the accumulation of cellular byproducts may inhibit cell growth, reduce antibody production, and adversely affect antibody glycosylation.

Amino acid concentration is also one of the important influencing factors; for example, in the culture medium, it affects cell growth and... IgG Productivity-related Leu and Arg The levels need to be well controlled. Amino acids with the highest consumption rates are associated with the production of... IgG The most abundant amino acids are closely associated with the process, so it is essential to ensure their sufficient availability during cultivation. Case studies are necessary for understanding the impact of medium and process optimization on glycosylation. It is important to gain insight into cell growth and cellular metabolism, IgG The interplay between synthesis and glycosylation, as well as how these factors vary at the metabolic level across different cell lines and culture medium compositions, will facilitate the optimization of biological processes and the development of culture media.

United States 908devices The company has launched two products highlighting the importance of media development for early-stage process development—one of which is an online biochemical analyzer designed specifically for the real-time monitoring of two key parameters: glucose and lactate. Maven Based on the technical principles of dialysis membranes and bio-(enzyme) sensors, the selective dialysis membrane ensures that the sterile environment inside the container does not contaminate the non-strictly sterile transport buffer system while allowing only small molecules such as glucose and lactic acid to pass through. These molecules are transported via the buffer solution to the measurement unit, achieving non-invasive analysis without the need for sample collection. Another key technological component—the measurement unit—employs a bio-(enzyme) sensor that exhibits high specificity toward the analyte and maintains effective measurement accuracy even at low concentrations. This approach is particularly well-suited for fermentation processes involving small volumes. / This sampling method, which does not cause sample loss, enables high-frequency sampling while simultaneously reducing the risk of contamination. Highest. 2 A measurement frequency of once per minute enables real-time monitoring of glucose and lactate levels, making it particularly suitable for early-stage process exploration and the study of glucose metabolism, thereby facilitating the development of strategies. fed-batch the feeding strategy or the implementation of automatic feeding.

Another product REBEL As a dedicated analyzer for culture medium components, it employs microfluidic chip technology. CE-MS Capillary electrophoresis - the principle of mass spectrometry coupling), helping R&D personnel break away from traditional analytical characterization processes (which involve numerous steps and are time-consuming), for... REBEL The sample requires only a microscale volume to complete multiple analyses. <30ul ), less than a single analysis 10 Can be completed in minutes >30 Ingredient composition (Figure) 2 The concentration analysis, operational procedures, and sample preparation processes are remarkably simple, offering both efficiency and cost-effectiveness compared to traditional methods.

Comprehensively understand cell growth and metabolism. IgG Titers and Glycosylation and fed-batch The dynamics of glucose and amino acid consumption are of great significance for understanding antibody production, optimizing processes, and improving antibody yield and quality.

Keywords:

[Literature Review] The Impact of Amino Acid and Glucose Metabolism in Fed-Batch Cultures of CHO Cells on Antibody Production and Glycosylation

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