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Solid-phase and bead-based cytokine immunoassay: a comparison.

机译:固相和基于珠的细胞因子免疫测定:比较。

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摘要

Cytokines and chemoattractive cytokines (chemokines) are present in a wide variety of body fluids such as plasma, cerebrospinal fluid, bronchoaveolar fluid, amniotic fluid, synovial fluid, middle ear effusion fluid, and urine. Cytokines can be detected using classical solid-phase sandwich immunoassays such as enzyme-linked immunosorbent assay (ELISA) or with a bead based multiplex immunoassay (MIA). The physical chemical properties of the different body fluids (such as pH and total protein content) differ, which may have an impact on the outcome of the cytokine assay. Both ELISA as well as MIA cytokine detection systems are constructed by sandwiching the protein of interest between a capture and reporter antibody. When the biological sample contains heterophilic antibodies (such as in patients with auto-immune diseases), these non-specific antibodies can cause false positive results. During pathological conditions, cytokines may be found over a wide concentration range; likewise have to cover this dynamic range in a similar fashion. The correct (statistical) analysis of standard curves and (multiplexed) data are critical for proper interpretation. Classical ELISA based cytokine assays are robust, easy to use and very well suited for measurement of single cytokines. Due to an increased interest in the integral approach to understand biological processes (the omics era), multiplex immunoassays for detection of cytokines and the interpretation of these assays are gaining popularity.
机译:细胞因子和化学吸引细胞因子(趋化因子)存在于多种体液中,例如血浆,脑脊液,支气管肺泡液,羊水,滑液,中耳积液和尿液。可以使用经典的固相夹心免疫测定法(例如酶联免疫吸附测定(ELISA))或基于微珠的多重免疫测定法(MIA)检测细胞因子。不同体液的物理化学性质(例如pH和总蛋白质含量)不同,这可能对细胞因子测定的结果产生影响。 ELISA和MIA细胞因子检测系统都是通过将目标蛋白夹在捕获抗体和报告抗体之间来构建的。当生物样品中含有嗜异性抗体时(例如患有自身免疫性疾病的患者),这些非特异性抗体会导致假阳性结果。在病理状态下,细胞因子的浓度范围很广。同样必须以类似的方式覆盖此动态范围。标准曲线和(多重)数据的正确(统计)分析对于正确的解释至关重要。基于经典ELISA的细胞因子检测方法功能强大,易于使用,非常适合用于测量单种细胞因子。由于对理解生物学过程的整体方法的兴趣日益增加(组学时代),用于检测细胞因子的多重免疫测定法和这些测定法的解释正逐渐普及。

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