Highly concentrated cell culture media formulations are essential to develop next generation bioprocesses used to produce therapeutic monoclonal antibodies, fusion proteins, bispecific molecules and mAb fragments. Although cysteine/cystine and tyrosine are the first components preventing the development of highly concentrated complex cell culture media, leucine and isoleucine were identified as the next critical amino acids due to their limited solubility. This work sought to investigate highly soluble and readily bioavailable derivatives of both amino acids that may be used in batch, fed-batch or perfusion processes. The α-keto acids of Leu and Ile, keto leucine and keto isoleucine sodium salts, were tested in cell culture media and proved to be beneficial to increase the overall solubility of cell culture media formulations. These keto acids were readily bioavailable for various CHO cells and can be used in both media and feeds. The quality of the final recombinant protein was studied in processes using the precursors and the mechanism of amination was investigated in CHO cells. Altogether, both keto acids represent an alternative to their respective amino acids to develop highly concentrated cell culture media formulations to support next generation bioprocesses.

高浓度的细胞培养基配方对于开发用于生产治疗性单克隆抗体，融合蛋白，双特异性分子和mAb片段的下一代生物工艺至关重要。尽管半胱氨酸/胱氨酸和酪氨酸是阻止高浓度复杂细胞培养基发展的首个成分，但是由于其溶解度有限，亮氨酸和异亮氨酸被认为是下一个关键氨基酸。这项工作试图研究可用于分批，补料分批或灌注过程的两种氨基酸的高度可溶且易于生物利用的衍生物。在细胞培养基中测试了Leu和Ile的α-酮酸，酮亮氨酸和酮异亮氨酸钠盐，证明对增加细胞培养基配方的整体溶解度有益。这些酮酸易于被各种CHO细胞生物利用，可用于培养基和饲料。在使用前体的过程中研究了最终重组蛋白的质量，并在CHO细胞中研究了胺化的机理。总之，两种酮酸都代表了它们各自氨基酸的替代品，以开发高度浓缩的细胞培养基配方，以支持下一代生物过程。