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相关概念视频

Hybridoma Technology01:31

Hybridoma Technology

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Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
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Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells
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Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells

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一个系统的方法来识别序列框架,减少mAb的生产在一个短暂的中国子卵巢细胞表达系统中.

Alana C Szkodny1, Kelvin H Lee1

  • 1Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.

Biotechnology progress
|April 12, 2024
PubMed
概括

抗体序列直接影响蛋白质表达水平. 这项研究确定了在CHO细胞中降低抗体表达的特定突变,有助于早期选择更好的候选药物.

科学领域:

  • 生物技术是生物技术.
  • 蛋白质工程是指蛋白质工程.
  • 生物制药制造业 生物制药制造业

背景情况:

  • 单克隆抗体 (mAbs) 是为了临床疗效而设计的,但在候选人选择之前,它们的"可开发性" (例如表达水平) 往往被忽视.
  • 与序列变化相关的表达不良,需要昂贵的制造工艺改进.
  • 难以表达的 (DTE) mAbs在折叠,组装和分泌方面表现出翻译后瓶,但发现很难泛化.

研究的目的:

  • 系统地研究序列变化的影响mAb表达在工业相关条件下的规模.
  • 为了确定影响中国仓鼠卵巢 (CHO) 细胞mAb表达的特定序列决定因素.
  • 开发一个工作流程,以改善mAb候选人选择和缩短流程开发时间表.

主要方法:

  • 设计了一个具有序列修改的单克隆抗体 (mAb) 变体库.
  • 这些变异的评估过渡表达标位在中国仓鼠卵巢 (CHO) 细胞中.
  • 分析了特定突变,残留性质 (如疏水性) 和表达水平之间的关系.

主要成果:

  • 确定了91种突变,显著降低了IgG1κ抗体的短暂表达.
  • 发现无法获得的残留物中的突变,特别是那些降低疏水性的人,对高抗体表达有害.
关键词:
在CHO细胞中.开发能力 开发能力难以表达的难以表达的一个单克隆抗体.短暂的表达方式 短暂的表达方式

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  • 在初级氨基酸序列和抗体表达标位之间显示出强烈的相关性,其中变异导致高达十倍的差异.

    • 结论:

    • 序列变化显著影响mAb表达,呈现出一个关键的可开发性特征.
    • 了解序列表达关系允许早期识别有问题的mAbs.
    • 这种系统的方法增强了抗体候选人的选择,并简化了生物过程的开发.