Aptamer as a Versatile Molecular Tool for Antibody Production Monitoring and Quality Control,Journal of the American Chemical Society

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Aptamer as a Versatile Molecular Tool for Antibody Production Monitoring and Quality Control
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-06-09 , DOI: 10.1021/jacs.9b13370
Kaiming Chen ₁ , Jie Zhou ₂ , Zhentao Shao ₁ , Jia Liu ₁ , Jia Song ₁ , Ruowen Wang ₁ , Juan Li _{1,

2} , Weihong Tan _{1,

3}

Affiliation

Antibody drugs have been used to treat many diseases, and to date, this has been the most rapidly growing drug class. However, the lack of suitable methods for real-time and high-throughput monitoring of antibody production and quality control has been a hindrance to further advancement of antibody drugs or biosimilars. Therefore, we herein report a versatile tool for one-step fluorescence monitoring of antibody production by using aptamer probes selected through the in vitro SELEX method. In this case, DNA aptamers were selected against the humanized IgG1 antibody drug trastuzumab with high specificity and affinity with a Kd value of aptamer CH1S-3 of 10.3 nM. More importantly, the obtained aptamers were able to distinguish native from heat-treated, whereas antibodies failed this test. Based on the advantages of rapid detection for aptamer, we designed aptamer molecular beacons for direct and sensitive detection of trastuzumab in complex samples. Unlike traditional antibody-based ELISA, the signal was observed directly upon interaction with target without the need for time-consuming binding and multiple washing steps. To further highlight biomedical applications, the use of aptamers as potential tools for quality control and traceless purification of antibody drugs was also demonstrated. Thus, aptamers are shown to be promising alternatives for antibody production monitoring, quality control and purification, providing technical support to accelerate antibody drug development.

中文翻译：

适体作为抗体生产监测和质量控制的多功能分子工具

抗体药物已被用于治疗许多疾病，迄今为止，这是增长最快的药物类别。然而，缺乏对抗体生产和质量控制进行实时和高通量监测的合适方法已经阻碍了抗体药物或生物仿制药的进一步发展。因此，我们在此报告了一种通用工具，通过使用通过体外 SELEX 方法选择的适体探针，对抗体生产进行一步荧光监测。在这种情况下，针对人源化 IgG1 抗体药物曲妥珠单抗选择了具有高特异性和亲和力的 DNA 适配体，适配体 CH1S-3 的 Kd 值为 10.3 nM。更重要的是，获得的适配体能够区分天然和热处理，而抗体未能通过该测试。基于适体快速检测的优势，我们设计了适体分子信标，用于直接和灵敏地检测复杂样品中的曲妥珠单抗。与传统的基于抗体的 ELISA 不同，信号在与靶标相互作用时直接观察到，无需耗时的结合和多次洗涤步骤。为了进一步突出生物医学应用，还证明了使用适体作为质量控制和无痕纯化抗体药物的潜在工具。因此，适配体被证明是抗体生产监测、质量控制和纯化的有前途的替代品，为加速抗体药物开发提供技术支持。在与目标相互作用时直接观察到信号，无需耗时的结合和多次洗涤步骤。为了进一步突出生物医学应用，还证明了使用适体作为质量控制和无痕纯化抗体药物的潜在工具。因此，适配体被证明是抗体生产监测、质量控制和纯化的有前途的替代品，为加速抗体药物开发提供技术支持。在与目标相互作用时直接观察到信号，无需耗时的结合和多次洗涤步骤。为了进一步突出生物医学应用，还证明了使用适体作为质量控制和无痕纯化抗体药物的潜在工具。因此，适配体被证明是抗体生产监测、质量控制和纯化的有前途的替代品，为加速抗体药物开发提供技术支持。

更新日期：2020-06-09

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