Merck Life Science, Upstream R&D, Darmstadt, Germany.
Merck Life Science, Process Development, Buchs, Switzerland.
Biotechnol Bioeng. 2021 Sep;118(9):3395-3408. doi: 10.1002/bit.27755. Epub 2021 Apr 8.
Increasing demands for protein-based therapeutics such as monoclonal antibodies, fusion proteins, bispecific molecules, and antibody fragments require researchers to constantly find innovative solutions. To increase yields and decrease costs of next generation bioprocesses, highly concentrated cell culture media formulations are developed but often limited by the low solubility of amino acids such as tyrosine, cystine, leucine, and isoleucine, in particular at physiological pH. This study sought to investigate highly soluble and bioavailable derivatives of leucine and isoleucine that are applicable for fed-batch processes. N-lactoyl-leucine and N-lactoyl-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 modified amino acids proved to be bioavailable for various Chinese hamster ovary (CHO) cells and were suitable for replacement of canonical amino acids in cell culture feeds. The quality of the final recombinant protein was studied in bioprocesses using the derivatives, and the mechanism of cleavage was investigated in CHO cells. Altogether, both N-lactoyl amino acids represent an advantageous alternative to canonical amino acids to develop highly concentrated cell culture media formulations to support next generation bioprocesses.
随着对蛋白质类治疗药物（如单克隆抗体、融合蛋白、双特异性分子和抗体片段）的需求不断增加，研究人员需要不断寻找创新的解决方案。为了提高下一代生物工艺的产量并降低成本，人们开发了高浓度的细胞培养基配方，但这些配方常常受到一些氨基酸（如酪氨酸、半胱氨酸、亮氨酸和异亮氨酸）溶解度低的限制，特别是在生理 pH 值条件下。本研究旨在探索亮氨酸和异亮氨酸的高溶解性和生物可利用衍生物，这些衍生物可用于补料分批工艺。研究测试了 N-乳酰-亮氨酸和 N-乳酰-异亮氨酸钠盐在细胞培养基中的应用，结果表明这两种盐有利于提高细胞培养基配方的总体溶解性。这些经过修饰的氨基酸被证明对各种中国仓鼠卵巢（CHO）细胞具有生物利用度，并且可替代细胞培养基补料中的常见氨基酸。研究人员在生物工艺中使用这些衍生物来研究最终重组蛋白的质量，并在 CHO 细胞中研究了裂解的机制。总之，N-乳酰氨基酸是替代常见氨基酸的有利选择，可以开发支持下一代生物工艺的高浓度细胞培养基配方。
