Kaissarian Nayiri M, Sandefur Stephanie L, Ghosh Arnab, Katneni Upendra K, Walton Wendy, Frye Christopher C, Komar Anton A, Kimchi-Sarfaty Chava
Hemostasis Branch 1, Division of Hemostasis, Office of Plasma Protein Therapeutics, Office of Therapeutic Products Center for Biologics Evaluation & Research, US Food and Drug Administration Silver Spring Maryland USA.
Bioproduct Research and Development Eli Lilly and Company Indiana USA.
Bioeng Transl Med. 2025 Jan 27;10(3):e10750. doi: 10.1002/btm2.10750. eCollection 2025 May.
Monoclonal antibodies (mAbs) are commonly used biologic therapeutics with a wide variety of clinical applications. During the development process, manufacturers consider different production parameters to improve protein yield and achieve appropriate quality of the product. Synonymous gene recoding is one of such attributes that is often considered and implemented to enhance protein expression. However, it has to be used with caution, as it may lead to protein misfolding and ER stress, which complicates efforts to manufacture the desired mAb. To investigate how changing mRNA sequence composition under different protein production parameters might affect the quality of recombinantly produced mAbs, we performed a comprehensive and systematic study assessing impact of synonymous gene recoding (commonly referred to as codon optimization) strategies in the context of varied cell culture parameters on product quality, biochemical and functional characteristics. We report the impact of these parameters on mAb glycosylation profiles, charge variant profile, aggregation, fragmentation, and mAb functional response from combinations of different production parameters. These results uncovered a complex interplay of sequence composition and manufacturing parameters and emphasize the importance of assessing changes to key quality attributes when optimizing mAb manufacturing, including the use of synonymous gene recoding.
单克隆抗体（mAb）是常用的生物治疗药物，具有广泛的临床应用。在开发过程中，制造商考虑不同的生产参数以提高蛋白质产量并实现产品的适当质量。同义基因编码是经常被考虑和实施以增强蛋白质表达的此类属性之一。然而，必须谨慎使用，因为它可能导致蛋白质错误折叠和内质网应激，这会使生产所需单克隆抗体的工作变得复杂。为了研究在不同蛋白质生产参数下改变mRNA序列组成如何影响重组生产的单克隆抗体的质量，我们进行了一项全面而系统的研究，评估了在不同细胞培养参数背景下同义基因编码（通常称为密码子优化）策略对产品质量、生化和功能特性的影响。我们报告了这些参数对单克隆抗体糖基化谱、电荷变体谱、聚集、片段化以及不同生产参数组合下单克隆抗体功能反应的影响。这些结果揭示了序列组成和制造参数之间复杂的相互作用，并强调了在优化单克隆抗体制备时评估关键质量属性变化的重要性，包括使用同义基因编码。
