Modulating antibody N-glycosylation through feed additives using a multi-tiered approach

IF 4.3 3区工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Frontiers in Bioengineering and Biotechnology Pub Date : 2024-08-26 DOI:10.3389/fbioe.2024.1448925

Jaka Kranjc, Lovro Kramer, Miha Mikelj, Marko Anderluh, Anja Pišlar, Matjaž Brinc

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引用次数: 0

Abstract

Glycosylation of recombinant proteins is a post-translational modification that affects multiple physicochemical and biological properties of proteins. As such, it is a critical quality attribute that must be carefully controlled during protein production in the pharmaceutical industry. Glycosylation can be modulated by various conditions, including the composition of production media and feeds. In this study, the N-glycosylation-modulating effects of numerous compounds, including metal enzyme cofactors, enzyme inhibitors, and metabolic intermediates, were evaluated. Chinese hamster ovary cells producing three different IgG antibodies were cultivated in a fed-batch mode. First, a one-factor-at-a-time experiment was performed in 24-well deep well plates to identify the strongest modulators and appropriate concentration ranges. Then, a full response surface experiment was designed to gauge the effects and interactions of the 14 most effective hit compounds in an Ambr® 15 bioreactor system. A wide range of glycoform content was achieved, with an up to eight-fold increase in individual glycoforms compared to controls. The resulting model can be used to determine modulator combinations that will yield desired glycoforms in the final product.

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采用多层次方法，通过饲料添加剂调节抗体 N-糖基化

重组蛋白质的糖基化是一种翻译后修饰，会影响蛋白质的多种物理化学和生物学特性。因此，糖基化是制药业蛋白质生产过程中必须谨慎控制的关键质量属性。糖基化可受各种条件的影响，包括生产介质和饲料的成分。本研究评估了多种化合物（包括金属酶辅助因子、酶抑制剂和代谢中间产物）对 N-糖基化的调节作用。以喂养批次模式培养了产生三种不同 IgG 抗体的中国仓鼠卵巢细胞。首先，在 24 孔深孔板中进行了一次一因素实验，以确定最强的调节剂和适当的浓度范围。然后，在 Ambr® 15 生物反应器系统中设计了一个完整的响应面实验，以衡量 14 种最有效的命中化合物的效果和相互作用。与对照组相比，单个糖形的含量最多可增加 8 倍。由此产生的模型可用于确定能在最终产品中产生理想糖型的调节剂组合。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Frontiers in Bioengineering and Biotechnology Chemical Engineering-Bioengineering

CiteScore

8.30

自引率

5.30%

发文量

2270

审稿时长

12 weeks

期刊介绍： The translation of new discoveries in medicine to clinical routine has never been easy. During the second half of the last century, thanks to the progress in chemistry, biochemistry and pharmacology, we have seen the development and the application of a large number of drugs and devices aimed at the treatment of symptoms, blocking unwanted pathways and, in the case of infectious diseases, fighting the micro-organisms responsible. However, we are facing, today, a dramatic change in the therapeutic approach to pathologies and diseases. Indeed, the challenge of the present and the next decade is to fully restore the physiological status of the diseased organism and to completely regenerate tissue and organs when they are so seriously affected that treatments cannot be limited to the repression of symptoms or to the repair of damage. This is being made possible thanks to the major developments made in basic cell and molecular biology, including stem cell science, growth factor delivery, gene isolation and transfection, the advances in bioengineering and nanotechnology, including development of new biomaterials, biofabrication technologies and use of bioreactors, and the big improvements in diagnostic tools and imaging of cells, tissues and organs. In today`s world, an enhancement of communication between multidisciplinary experts, together with the promotion of joint projects and close collaborations among scientists, engineers, industry people, regulatory agencies and physicians are absolute requirements for the success of any attempt to develop and clinically apply a new biological therapy or an innovative device involving the collective use of biomaterials, cells and/or bioactive molecules. “Frontiers in Bioengineering and Biotechnology” aspires to be a forum for all people involved in the process by bridging the gap too often existing between a discovery in the basic sciences and its clinical application.

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