论文部分内容阅读
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。
OBJECTIVE: To explore the immunoreaction process of magnetic beads at the microscopic level by exploring the effective volume of immunoreaction (NEIV) of single magnetic beads and provide theoretical guidance for the amount of magnetic beads used in the detection of large volume samples. Innovative point: For the first time, we proposed the concept of “effective volume of immunoreaction per single bead”, and put forward to probe into the immunological reaction of beads at the microscopic level, which laid the foundation for further study on the kinetics of bead-antigen immunoreactions. Methods: First, colorimetric detection of 50 μl of different concentrations of antigen using two diameters of magnetic beads demonstrated that the antigen binding sites on the beads were excessive. Subsequently, quantitative magnetic beads were used to detect the different concentrations of antigens of the same concentration to arrive at the concept of “effective immunoreaction volume per single bead”. Next, three different amounts of magnetic beads were used to test for antigens of the same concentration and increasing volume gradients to validate the proposed concept of NEIV. Then, find out the key factors affecting the NEIV by measuring the effect of incubation time, incubation temperature and shock on the colorimetric results. Finally, quantitative magnetic beads were used to detect the gradient-diluted antigen. The relationship between the amount of magnetic beads and the colorimetric results was established, and the prospect of NEIV was put forward. Conclusion: For the same diameter of magnetic beads, the NEIV of a single magnetic bead is constant under the same incubation conditions, regardless of the amount of magnetic beads and the incubation volume. Incubation temperature, incubation duration and shock or not are the key factors affecting the NEIV value. Using the NEIV parameter provides an effective guide to the bead usage simulation prior to large sample size testing. In addition, NEIV revealed the immunological reactions of magnetic beads and antigens at the microscopic level, which laid the foundation for further study on the kinetics of magnetic bead-antigen immunoreactions.