Buffer system improves the removal of host cell protein impurities in monoclonal antibody purification

IF 3.5 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Dániel Lakatos, Martina Idler, Selina Stibitzky, Jennifer Amann, Jakob Schuschkewitz, Dominik Krayl, Judith Liebau, Jan-Hendrik Grosch, Erik Arango Gutierrez, Simon Kluters
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引用次数: 0

Abstract

Polysorbates (PS) are commonly used as stabilizers of biopharmaceuticals such as monoclonal antibodies (mAbs). However, they are prone to chemical and enzymatic degradation. The latter can be caused by residual host cell proteins (HCPs) in the drug substance. Degradation affects the functionality of the PS surfactant which can lead to formation of particles. An increasing number of publications describe enzymatic PS degradation. Significant efforts have been made to characterize HCP removal during Downstream Processing (DSP) of mAbs and to develop mitigation strategies. Here we describe the use of glycine buffer for acidic elution in Protein A affinity chromatography compared to acetate buffer, which is more commonly used in the biopharmaceutical industry. Increased turbidity was observed during pH re-adjustment after low pH virus inactivation when using glycine buffer. Analytical data suggests that this turbidity is caused by the formation of precipitates which include HCP and DNA impurities. Additionally, as a zwitterion, glycine does not contribute to conductivity; this further enhances HCP removal during anion-exchange flow-through chromatography. Although glycine is well known as a possible elution buffer for Protein A affinity chromatography, its positive impact on HCP removal and PS stability have not yet been described in literature.
缓冲液系统可改善单克隆抗体纯化过程中宿主细胞蛋白质杂质的去除效果
聚山梨醇酯(PS)通常用作单克隆抗体(mAbs)等生物制药的稳定剂。然而,它们容易发生化学和酶降解。后者可由药物中残留的宿主细胞蛋白(HCPs)引起。降解会影响 PS 表面活性剂的功能,从而导致颗粒的形成。越来越多的出版物对 PS 的酶降解进行了描述。为了描述 mAbs 下游加工 (DSP) 过程中 HCP 的去除情况并制定缓解策略,人们付出了巨大的努力。与生物制药行业更常用的醋酸盐缓冲液相比,我们在此介绍在蛋白 A 亲和层析中使用甘氨酸缓冲液进行酸性洗脱的情况。在使用甘氨酸缓冲液进行低 pH 值病毒灭活后的 pH 值再调整过程中,观察到浑浊度增加。分析数据表明,这种浑浊是由包括 HCP 和 DNA 杂质在内的沉淀物形成造成的。此外,作为一种齐聚物,甘氨酸不会对电导率产生影响;这进一步提高了阴离子交换流动色谱法对 HCP 的去除率。虽然甘氨酸作为蛋白质 A 亲和层析的洗脱缓冲液已广为人知,但其对 HCP 去除和 PS 稳定性的积极影响尚未在文献中有所描述。
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来源期刊
Biotechnology and Bioengineering
Biotechnology and Bioengineering 工程技术-生物工程与应用微生物
CiteScore
7.90
自引率
5.30%
发文量
280
审稿时长
2.1 months
期刊介绍: Biotechnology & Bioengineering publishes Perspectives, Articles, Reviews, Mini-Reviews, and Communications to the Editor that embrace all aspects of biotechnology. These include: -Enzyme systems and their applications, including enzyme reactors, purification, and applied aspects of protein engineering -Animal-cell biotechnology, including media development -Applied aspects of cellular physiology, metabolism, and energetics -Biocatalysis and applied enzymology, including enzyme reactors, protein engineering, and nanobiotechnology -Biothermodynamics -Biofuels, including biomass and renewable resource engineering -Biomaterials, including delivery systems and materials for tissue engineering -Bioprocess engineering, including kinetics and modeling of biological systems, transport phenomena in bioreactors, bioreactor design, monitoring, and control -Biosensors and instrumentation -Computational and systems biology, including bioinformatics and genomic/proteomic studies -Environmental biotechnology, including biofilms, algal systems, and bioremediation -Metabolic and cellular engineering -Plant-cell biotechnology -Spectroscopic and other analytical techniques for biotechnological applications -Synthetic biology -Tissue engineering, stem-cell bioengineering, regenerative medicine, gene therapy and delivery systems The editors will consider papers for publication based on novelty, their immediate or future impact on biotechnological processes, and their contribution to the advancement of biochemical engineering science. Submission of papers dealing with routine aspects of bioprocessing, description of established equipment, and routine applications of established methodologies (e.g., control strategies, modeling, experimental methods) is discouraged. Theoretical papers will be judged based on the novelty of the approach and their potential impact, or on their novel capability to predict and elucidate experimental observations.
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