本文關(guān)鍵詞： 單個磁珠有效免疫反應(yīng)體積(NEIV) 免疫磁珠 酶聯(lián)免疫吸附測定(ELISA) 磁珠直徑 孵育方式　出處：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2017年10期 　論文類型：期刊論文

【摘要】：目的:通過探究單個磁珠的有效免疫反應(yīng)體積(NEIV),在微觀水平上揭示磁珠的免疫反應(yīng)過程,為大體積樣本檢測過程中磁珠的用量提供理論指導(dǎo)。創(chuàng)新點:首次提出"單個磁珠的有效免疫反應(yīng)體積"這一概念,提出在微觀水平上對磁珠免疫反應(yīng)進行探究,為進一步研究磁珠-抗原免疫反應(yīng)的動力學(xué)過程打下基礎(chǔ)。方法:首先,采用兩種直徑的磁珠對50μl不同濃度的抗原進行比色檢測,證明磁珠上抗原的結(jié)合位點過量。隨后,采用定量磁珠對相同濃度不同體積的抗原進行檢測,得出"單個磁珠的有效免疫反應(yīng)體積"這一概念。接著,采用三種不同用量的磁珠對濃度相同、體積梯度增大的抗原進行檢測,以驗證所提出的NEIV的概念。然后,通過測定孵育時長、孵育溫度和震蕩與否對比色結(jié)果的影響,找到對NEIV影響的關(guān)鍵因素。最后采用定量磁珠,對梯度稀釋的抗原進行檢測,建立磁珠用量與比色結(jié)果的關(guān)系,對NEIV的實際應(yīng)用提出展望。結(jié)論:對于同一直徑的磁珠,在相同的孵育條件下,其單個磁珠的NEIV是恒定的,與磁珠用量和孵育體積無關(guān)。孵育溫度、孵育時長和震蕩與否是影響NEIV值的關(guān)鍵因素。采用NEIV這一參數(shù)可以在大體積樣本檢測之前,對磁珠的用量模擬提供有效指導(dǎo)。除此之外,NEIV更從微觀角度上揭示了磁珠和抗原的免疫反應(yīng)過程,為進一步研究磁珠-抗原免疫反應(yīng)的動力學(xué)過程奠定了基礎(chǔ)。
[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.
【作者單位】： 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
【分類號】：R446.6

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