Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems)
Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems)
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DOI：
10.1016/j.bbrc.2006.11.140
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发表时间：
2007-01-26
影响因子：
3.1
通讯作者：
Kovacs, Joseph A.
中科院分区：
文献类型：
--
作者：
Burbelo, Peter D.;Ching, Kathryn H.;Kovacs, Joseph A.
应用LIPS的快速定量抗体反应传染性病原体的描述。在哺乳动物细胞中表达与海肾荧光素酶融合的编码病原体抗原的嵌合基因;制备粗提物，并且不经纯化，用于免疫沉淀测定以定量病原体特异性抗体。在横断面和纵向研究中，通过该测定法测量的耶氏肺孢子虫MSG-14抗原的抗体水平与先前用优化的ELISA获得的水平相关性良好。我们还正确预测了B型肝炎（HBV）、丙型肝炎（HCV）和HIV感染状态，分析了33例患者血清的99次检测中只有2次除外。然后，我们使用了15个HIV编码的蛋白质，包括整个HIV蛋白质组，以产生抗体反应谱这33个血清。每种HIV抗原都能被至少一个HIV感染者血清中的抗体识别。用这些简单的定量抗体检测分析产生的数据具有临床和研究应用。(c)2006年爱思唯尔公司All rights reserved.
The application of LIPS to the rapid quantification of antibody responses to infectious agents is described. Chimeric genes encoding pathogen antigens fused to Renilla luciferase are expressed in mammalian cells; crude extracts are prepared and, without purification, employed in immunoprecipitation assays to quantify pathogen-specific antibodies. In cross-sectional and longitudinal studies, antibody levels to the MSG-14 antigen of Pneumocystis jirovecii measured by this assay correlated well with levels previously obtained with an optimized ELISA. We also correctly predicted Hepatitis B (HBV), Hepatitis C (HCV), and HIV infection status in all but 2 of 99 assays analyzing 33 patient sera. We then used 15 HIV-encoded proteins comprising the whole HIV proteome to generate antibody response profiles for these 33 sera. Each HIV antigen was recognized by antibodies in serum from at least one HIV-infected individual. Data generated with these simple, quantitative antibody-detection assays have both clinical and research applications. (c) 2006 Elsevier Inc. All rights reserved.