School of Interdisciplinary Research, Indian Institute of Technology Delhi, New Delhi, India.
Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
J Chromatogr A. 2024 Jan 25;1715:464605. doi: 10.1016/j.chroma.2023.464605. Epub 2023 Dec 23.
Monoclonal antibody downstream processing typically entails chromatography-based purification processes beginning with Protein A chromatography, accounting for 50 % of the total manufacturing expense. Alternatives to protein A chromatography have been explored by several researchers. In this paper, aqueous two-phase extraction (ATPE) has been proposed for continuous processing of monoclonal antibodies (mAbs) as an alternative to the traditional protein A chromatography. The PEG-sulfate system has been employed for phase formation in ATPE, and the mAb is separated in the salt phase, while impurities like high molecular weight (HMW) and host cell proteins (HCPs) are separated in the PEG phase. Following ATPE of clarified cell culture harvest, yield of ≥ 80 % and purity of ≥ 97 % were achieved in the salt phase. Considerable (28 %) reduction in consumable cost has been estimated when comparing the proposed platform to the traditional protein A based platform. The outcomes demonstrate that ATPE can be a potentially effective substitute for the traditional Protein A chromatography for purification of mAbs. The proposed platform offers easy implementation, delivers comparative results, and offers significantly better economics for manufacturing mAb-based biotherapeutics.
单克隆抗体下游处理通常需要基于色谱的纯化过程，从 Protein A 色谱法开始，占总制造成本的 50%。一些研究人员已经探索了替代 Protein A 色谱法的方法。在本文中，提出了用于单克隆抗体 (mAb) 连续处理的双水相萃取 (ATPE) 作为传统 Protein A 色谱法的替代方法。PEG-硫酸盐系统已用于 ATPE 中的相形成，mAb 分离在盐相中，而杂质如高分子量 (HMW) 和宿主细胞蛋白 (HCP) 则分离在 PEG 相中。在澄清的细胞培养物收获物进行 ATPE 后，在盐相中实现了≥80%的收率和≥97%的纯度。与传统的基于 Protein A 的平台相比，估计当比较所提出的平台时，消耗品成本可降低 28%。结果表明，ATPE 可以成为 mAb 纯化的传统 Protein A 色谱法的潜在有效替代品。所提出的平台易于实施，提供可比的结果，并为制造基于 mAb 的生物治疗药物提供了显著更好的经济性。
