本文關(guān)鍵詞： 單個(gè)磁珠有效免疫反應(yīng)體積(NEIV) 免疫磁珠 酶聯(lián)免疫吸附測(cè)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(lèi)型：期刊論文

【摘要】：目的:通過(guò)探究單個(gè)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(guò)程,為大體積樣本檢測(cè)過(guò)程中磁珠的用量提供理論指導(dǎo)。創(chuàng)新點(diǎn):首次提出"單個(gè)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(duì)磁珠免疫反應(yīng)進(jìn)行探究,為進(jìn)一步研究磁珠-抗原免疫反應(yīng)的動(dòng)力學(xué)過(guò)程打下基礎(chǔ)。方法:首先,采用兩種直徑的磁珠對(duì)50μl不同濃度的抗原進(jìn)行比色檢測(cè),證明磁珠上抗原的結(jié)合位點(diǎn)過(guò)量。隨后,采用定量磁珠對(duì)相同濃度不同體積的抗原進(jìn)行檢測(cè),得出"單個(gè)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(duì)濃度相同、體積梯度增大的抗原進(jìn)行檢測(cè),以驗(yàn)證所提出的NEIV的概念。然后,通過(guò)測(cè)定孵育時(shí)長(zhǎng)、孵育溫度和震蕩與否對(duì)比色結(jié)果的影響,找到對(duì)NEIV影響的關(guān)鍵因素。最后采用定量磁珠,對(duì)梯度稀釋的抗原進(jìn)行檢測(cè),建立磁珠用量與比色結(jié)果的關(guān)系,對(duì)NEIV的實(shí)際應(yīng)用提出展望。結(jié)論:對(duì)于同一直徑的磁珠,在相同的孵育條件下,其單個(gè)磁珠的NEIV是恒定的,與磁珠用量和孵育體積無(wú)關(guān)。孵育溫度、孵育時(shí)長(zhǎng)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(cè)之前,對(duì)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(guò)程,為進(jìn)一步研究磁珠-抗原免疫反應(yīng)的動(dòng)力學(xué)過(guò)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(lèi)號(hào)】：R446.6

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