R&D Laboratory, Diagnosticum Zrt, 1047 Budapest, Hungary.
Department of Applied Analysis and Computational Mathematics, Eötvös Loránd University, 1117 Budapest, Hungary.
Sensors (Basel). 2022 May 23;22(10):3962. doi: 10.3390/s22103962.
In spite of its pivotal role in the characterization of humoral immunity, there is no accepted method for the absolute quantitation of antigen-specific serum antibodies. We devised a novel method to quantify polyclonal antibody reactivity, which exploits protein microspot assays and employs a novel analytical approach. Microarrays with a density series of disease-specific antigens were treated with different serum dilutions and developed for IgG and IgA binding. By fitting the binding data of both dilution series to a product of two generalized logistic functions, we obtained estimates of antibody reactivity of two immunoglobulin classes simultaneously. These estimates are the antigen concentrations required for reaching the inflection point of thermodynamic activity coefficient of antibodies and the limiting activity coefficient of antigen. By providing universal chemical units, this approach may improve the standardization of serological testing, the quality control of antibodies and the quantitative mapping of the antibody-antigen interaction space.
尽管其在体液免疫特性描述中起着关键作用，但目前还没有公认的方法可以对抗原特异性血清抗体进行绝对定量。我们设计了一种新的方法来定量分析多克隆抗体反应，该方法利用了蛋白质微点分析，并采用了一种新的分析方法。用一系列具有不同密度的疾病特异性抗原处理微阵列，并用于 IgG 和 IgA 结合的研究。通过将两个稀释系列的结合数据拟合到两个广义逻辑函数的乘积中，我们同时获得了两种免疫球蛋白类别的抗体反应性估计值。这些估计值是达到抗体热力学活度系数拐点和抗原极限活度系数所需的抗原浓度。通过提供通用的化学单位，这种方法可能会提高血清学检测的标准化程度、抗体的质量控制以及抗体-抗原相互作用空间的定量映射。
