利用离子交换周期性逆流色谱法设计和优化低滴度酶的连续纯化工艺

IF 2.5 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology and Bioprocess Engineering Pub Date : 2024-04-04 DOI:10.1007/s12257-024-00099-1
Kwanyoung Ko, Min-Jung Kim, Dasom Kim, Kangyun Seo, Sangho Lee
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引用次数: 0

摘要

与传统的分批纯化工艺相比,连续纯化工艺的生产率和效率更高,因此有利于生物制剂的纯化。然而,由于监管问题和缺乏成熟案例,连续工艺很难在工业环境中应用。在此,我们报告了一个案例研究,用于设计和优化先进的连续工艺,以纯化一种低滴度酶作为示范生物制剂。为了将生物制剂纯化中的传统批次工艺转换为先进的连续工艺,传统的单元操作(UOs),包括超滤/渗滤(UF/DF)和批次色谱,被先进的单元操作(UOs),如在线稀释调节(IDC)和周期性逆流色谱(PCC)所取代。将 UF/DF UO 改为 IDC UO,以调节 pH 值和电导率。IDC 中样品与调节缓冲液的混合比例是通过三种缓冲液的实验确定的。由于色谱柱中的 delta 压力小于 1.0 巴,因此利用色谱柱高度和样品装载停留时间这两个变量对 PCC 进行了优化。绘制了显示操作区域的曲线图,以有效控制 PCC。虽然 IDC 中的样品量增加了,但 PCC 具有互补优势,即纯化速度比间歇层析快。我们观察到,当采用先进的连续工艺纯化低滴度酶时,经济效益至少提高了 25%。我们不仅提出了一种用 IDC 和 PCC 替代 UF/DF 和批次色谱的连续工艺,还提出了一种通过绘制操作区来优化 PCC 的方法。
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Design and optimization of a continuous purification process using ion-exchange periodic counter-current chromatography for a low-titer enzyme

A continuous purification process can be beneficial to the purification of biologics due to its higher productivity and efficiency than a conventional batch purification process. However, regulatory issues and lack of established cases render deployment of the continuous process difficult in industrial settings. Here we report a case study for design and optimization of an advanced continuous process for purifying a low-titer enzyme as a model biologic. To convert a conventional batch process to an advanced continuous one in purification of biologics, conventional unit operations (UOs), including ultrafiltration/diafiltration (UF/DF) and batch chromatography, were replaced by advanced ones such as in-line dilution conditioning (IDC) and periodic counter-current chromatography (PCC). The UF/DF UO was changed to IDC UO to adjust pH and conductivity. The mixing ratio of the sample and the conditioning buffer in IDC was determined by experiments with three buffers. PCC was optimized with two variables, column height and sample loading residence time, as the delta pressure in the columns was less than 1.0 bar. A graph indicating the operating area was plotted to efficiently control the PCC. Although the sample volume increased in IDC, PCC had a complementary advantage in that purification was performed faster than batch chromatography. We observed at least 25% increase in economic advantage when the advanced continuous process was applied to purify a low-titer enzyme. We propose not only a continuous process with the substitution of UF/DF and batch chromatography with IDC and PCC but also a method to optimize PCC by plotting operating areas.

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来源期刊
Biotechnology and Bioprocess Engineering
Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物
CiteScore
5.00
自引率
12.50%
发文量
79
审稿时长
3 months
期刊介绍: Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.
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