在单个磁珠的水平上探究磁珠的有效免疫反应体积（英文）

发布时间：2018-02-13 15:31

本文关键词： 单个磁珠有效免疫反应体积(NEIV) 免疫磁珠酶联免疫吸附测定(ELISA) 磁珠直径孵育方式　出处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2017年10期 　论文类型：期刊论文

【摘要】：目的:通过探究单个磁珠的有效免疫反应体积(NEIV),在微观水平上揭示磁珠的免疫反应过程,为大体积样本检测过程中磁珠的用量提供理论指导。创新点:首次提出"单个磁珠的有效免疫反应体积"这一概念,提出在微观水平上对磁珠免疫反应进行探究,为进一步研究磁珠-抗原免疫反应的动力学过程打下基础。方法:首先,采用两种直径的磁珠对50μl不同浓度的抗原进行比色检测,证明磁珠上抗原的结合位点过量。随后,采用定量磁珠对相同浓度不同体积的抗原进行检测,得出"单个磁珠的有效免疫反应体积"这一概念。接着,采用三种不同用量的磁珠对浓度相同、体积梯度增大的抗原进行检测,以验证所提出的NEIV的概念。然后,通过测定孵育时长、孵育温度和震荡与否对比色结果的影响,找到对NEIV影响的关键因素。最后采用定量磁珠,对梯度稀释的抗原进行检测,建立磁珠用量与比色结果的关系,对NEIV的实际应用提出展望。结论:对于同一直径的磁珠,在相同的孵育条件下,其单个磁珠的NEIV是恒定的,与磁珠用量和孵育体积无关。孵育温度、孵育时长和震荡与否是影响NEIV值的关键因素。采用NEIV这一参数可以在大体积样本检测之前,对磁珠的用量模拟提供有效指导。除此之外,NEIV更从微观角度上揭示了磁珠和抗原的免疫反应过程,为进一步研究磁珠-抗原免疫反应的动力学过程奠定了基础。
[Abstract]:Objective: to investigate the effective immune response volume of single magnetic beads and reveal the immune response process of magnetic beads at microcosmic level. To provide theoretical guidance for the amount of magnetic beads in the detection process of large samples. Innovation: the concept of "effective immune response volume of single magnetic beads" is proposed for the first time, and the research on the immune response of magnetic beads at the micro level is put forward. Methods: firstly, two kinds of magnetic beads with diameter were used to detect 50 渭 l antigens of different concentration, which proved that the binding sites of antigens on magnetic beads were excessive. Using quantitative magnetic beads to detect antigens of the same concentration and different volume, the concept of "effective immune response volume of single magnetic beads" was obtained. Then, three kinds of magnetic beads with the same concentration were used. The antigen with increasing volume gradient was detected to verify the proposed concept of NEIV. Then, by measuring the effect of incubation time, incubation temperature and the contrast color result of shaking or not, the key factors affecting NEIV were found. Finally, quantitative magnetic beads were used. The relationship between the amount of magnetic beads and colorimetric results was established, and the practical application of NEIV was prospected. Conclusion: for magnetic beads of the same diameter, the NEIV of single magnetic beads is constant under the same incubation conditions. It has nothing to do with the amount of magnetic beads and the volume of incubation. Incubation temperature, incubation time and oscillation are the key factors that affect the NEIV value. The NEIV parameter can be used before large samples are tested. In addition, NEIV revealed the immunoreaction process of magnetic beads and antigens from the microscopic point of view, which laid a foundation for further study on the kinetic process of magnetic bead antigen immune reaction.
【作者单位】： College
【基金】：Project supported by the National Natural Science Foundation of China (No.31571918) Hong Kong, Macao, and Taiwan Scientific and Technological Cooperation Projects (No.2015DFT30150) of China the Zhejiang Provincial Department of Education (No.Y201533676), China
【分类号】：R446.6

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