Control and estimation of glycosylation profile via media supplementation based on intracellular models in mammalian cell cultures

基于哺乳动物细胞培养物中的细胞内模型通过培养基补充来控制和估计糖基化谱

• 批准号: 1706731
• 负责人: Seongkyu Yoon
• 金额: $ 32.95万
• 依托单位: University of Massachusetts Lowell
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1706731&HistoricalAwards=false
• 关键词: Control; estimation; glycosylation; profile; via

Recombinant monoclonal antibodies (mAbs) are effective in curing many diseases, including several types of cancer. The effectiveness of mAbs is closely related to the pattern of decoration of the mAb surface with sugars. This project attempts to understand and ultimately predict how mAbs are decorated with sugars in specific locations on the surface. Such knowledge would fill a significant gap in current industrial practice on mAb quality control and improve the development and manufacturing of such therapeutic molecules. The research team will also provide STEM training opportunities through undergraduate research, recruiting minority students, developing new biomanufacturing courses and lectures, and supporting outreach activities for K-12 students and general public. Successful control and optimization of mAb glycosylation is currently hindered by limited understanding of how glycosylation responds in vivo to culture perturbations. This research proposes to tackle the challenge by exploring intracellular metabolic mechanisms that link media supplementation in to glycosylation.This should eventually lead to the establishment of a model to predict glycosylation variation in cultures and accommodate a control methodology via media supplementation. Mechanistic insights into the cellular physiology critical to glycosylation will be achieved by multi-omics approaches. This will pave the way to developing intracellular metabolic models which connect media supplements, intracellular properties, and glycan profiles, an to developing control strategies for glycan attributes via media supplementation. If successful, these results should enable the reproducible generation of fully active monoclonal antibodies, increasing the efficacy and decreasing the cost of this important human health therapy.

重组单克隆抗体（mab）可有效治疗多种疾病，包括几种类型的癌症。单克隆抗体的有效性与单克隆抗体表面的糖修饰模式密切相关。这个项目试图理解并最终预测单克隆抗体是如何在表面的特定位置被糖修饰的。这些知识将填补当前工业实践中单克隆抗体质量控制的重大空白，并改善此类治疗分子的开发和制造。研究团队还将通过本科生研究、招募少数民族学生、开发新的生物制造课程和讲座，以及支持面向K-12学生和公众的外展活动，提供STEM培训机会。单抗糖基化的成功控制和优化目前受到对体内糖基化如何响应培养扰动的有限理解的阻碍。本研究提出通过探索将培养基补充与糖基化联系起来的细胞内代谢机制来解决这一挑战。这最终将导致建立一个模型来预测培养中的糖基化变化，并通过培养基补充适应控制方法。对糖基化至关重要的细胞生理学的机制见解将通过多组学方法实现。这将为建立细胞内代谢模型铺平道路，该模型将介质补充、细胞内特性和聚糖谱联系起来，并通过介质补充开发聚糖属性的控制策略。如果成功，这些结果将使完全活性单克隆抗体的可重复产生成为可能，从而提高这种重要的人类健康疗法的功效并降低其成本。

项目成果

An investigation of nucleotide sugar dynamics under the galactose supplementation in CHO cell culture

CHO 细胞培养物中半乳糖补充下核苷酸糖动态的研究

• DOI: 10.1016/j.procbio.2019.03.020
• 发表时间: 2019
• 期刊: Process Biochemistry
• 影响因子: 4.4
• 作者: Sha, Sha;Yoon, Seongkyu
• 通讯作者: Yoon, Seongkyu

Prediction of N-linked Glycoform Profiles of Monoclonal Antibody with Extracellular Metabolites and Two-Step Intracellular Models

使用细胞外代谢物和两步细胞内模型预测单克隆抗体的 N 连接糖型谱

• DOI: 10.3390/pr7040227
• 发表时间: 2019
• 期刊: Processes
• 影响因子: 3.5
• 作者: Sha, Sha;Huang, Zhuangrong;Agarabi, Cyrus;Lute, Scott;Brorson, Kurt;Yoon, Seongkyu
• 通讯作者: Yoon, Seongkyu

Mechanistic modeling and applications for CHO cell culture development and production

• DOI: 10.1016/j.coche.2018.08.010
• 发表时间: 2018-12
• 期刊: Current Opinion in Chemical Engineering
• 影响因子: 6.6
• 作者: Sha Sha-Sha;Zhuangrong Huang;Zhao Wang;Seongkyu Yoon

A high-resolution measurement of nucleotide sugars by using ion-pair reverse chromatography and tandem columns

• DOI: 10.1007/s00216-020-02608-6
• 发表时间: 2020-06-01
• 期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
• 影响因子: 4.3
• 作者: Sha, Sha;Handelman, Garry;Yoon, Seongkyu
• 通讯作者: Yoon, Seongkyu

At-line N-linked glycan profiling for monoclonal antibodies with advanced sample preparation and high-performance liquid chromatography

• DOI: 10.1016/j.jbiosc.2020.04.009
• 发表时间: 2020-09-01
• 期刊: JOURNAL OF BIOSCIENCE AND BIOENGINEERING
• 影响因子: 2.8
• 作者: Sha, Sha;Handelman, Garry;Yoon, Seongkyu
• 通讯作者: Yoon, Seongkyu