Antibody profiling by Luciferase Immunoprecipitation Systems (LIPS).
Antibody profiling by Luciferase Immunoprecipitation Systems (LIPS).
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DOI：
10.3791/1549
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发表时间：
2009-10-07
期刊：
影响因子：
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通讯作者：
Iadarola, Michael J
中科院分区：
文献类型：
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作者：
Burbelo, Peter D;Ching, Kathryn H;Iadarola, Michael J
用于全面理解和量化自身抗原和感染因子的抗体谱的技术可能会对疾病机制产生新的见解，并可能阐明新的标志物以替代具有不同临床特征的疾病并更好地理解发病机制。我们已经开发了一种称为荧光素酶免疫沉淀系统（LIPS）的高度定量方法，用于分析患者血清对与感染相关的自身抗原和病原体抗原的抗体反应。与ELISA不同，高灵敏度的LIPS易于实施，以通用格式调查对不同抗原的体液血清学应答谱，并对某些抗原产生高达6 log（10）的动态抗体滴度范围。在这些研究中，通过LIPS对患者针对抗原组或甚至某些病原体（即HIV）的整个蛋白质组的体液应答进行定量分析，通常比通过ELISA检测单个抗原提供更多信息。此外，LIPS还消除了生产高度纯化抗原所需的时间和精力，以及标准ELISA所需的劳动密集型测定优化步骤。在这里，我们提供了一个详细的协议，描述了技术方面的执行LIPS测定容易剖析抗体反应的单一或多种抗原。
Technologies for comprehensively understanding and quantifying antibody profiles to autoantigens and infectious agents may yield new insights into disease mechanisms and may elucidate new markers to substratify disease with different clinical features and better understand pathogenesis. We have developed a highly quantitative method called Luciferase Immunoprecipitation Systems (LIPS) for profiling patient sera antibody responses to autoantigens and pathogen antigens associated with infection. Unlike ELISAs, the highly sensitive LIPS is easily implemented to survey humoral serological response profiles to different antigens in a universal format and produces dynamic antibody titer ranges up to 6 log(10) for some antigens. In these studies, quantitative profiling by LIPS of patient humoral responses against panels of antigens or even the entire proteome of some pathogens (i.e. HIV), is typically more informative than testing a single antigen by ELISA. In addition, LIPS also eliminates time and effort needed to produce highly purified antigens as well as the labor-intensive assay optimization steps needed for standard ELISAs. Here we provide a detailed protocol describing the technical aspects of performing LIPS assays for readily profiling antibody responses to single or multiple antigens.