Development of a high-throughput bead based assay system to measure HIV-1 specific immune signatures in clinical samples
Development of a high-throughput bead based assay system to measure HIV-1 specific immune signatures in clinical samples
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DOI：
10.1016/j.jim.2017.12.003
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发表时间：
2018-03-01
影响因子：
2.2
通讯作者：
Trkola, Alexandra
中科院分区：
文献类型：
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作者：
Liechti, Thomas;Kadelka, Claus;Trkola, Alexandra
监测和评估基于广泛中和抗体（bnAb）的HIV-1疫苗需要详细测量HIV-1结合抗体应答，以支持检测保护相关性。在这里，我们描述了一个灵活的，高通量的微球为基础的多重检测系统，允许监测复杂的结合抗体的签名的发展。研究一组13种HIV-1抗原的不同IgG亚类（IgG 1，IgG 2和IgG 3）的平行评估，我们证明了我们的策略的潜力。我们描述的技术进步包括使用抗原的定向中性亲和素-生物素固定化和生物素饱和以减少背景来改善抗原反应性的手段。我们的研究的一个特别强调的是提供用于评估测定系统的再现性和稳定性的工具以及控制允许在高通量测定中应用的变化的策略，其中需要保证单次测量的可靠性。
The monitoring and assessment of a broadly neutralizing antibody (bnAb) based HIV-1 vaccine require detailed measurements of HIV-1 binding antibody responses to support the detection of correlates of protection. Here we describe the development of a flexible, high-throughput rnicrosphere based multiplex assay system that allows monitoring complex binding antibody signatures. Studying a panel of 13 HIV-1 antigens in a parallel assessment of different IgG subclasses (IgG1, IgG2 and IgG3) we demonstrate the potential of our strategy. The technical advances we describe include means to improve antigen reactivity using directed neutravidin-biotin immobilization of antigens and biotin saturation to reduce background. A particular emphasis of our study was to provide tools for the assessment of reproducibility and stability of the assay system and strategies to control for variations allowing the application in high throughput assays, where reliability of single measurements needs to be guaranteed.