A new approach to study drugs containing ultra-high dilutions of antibodies using fluorescence spectroscopy

This data set contains results of ultrahigh dilutions effects study. The physicochemical (and therefore, spectral) properties of lactose water solutions change in the presence of ultrahigh dilutions of antibodies (UHD Ab). Spectrofluorimetric assessment of solutions containing UHD Ab to interferon gamma (IFNg)were performed and quantitative differences (420 nm peak emission intensity) from control samples were determined . The validation study of the procedure has shown that the proposed method is specific: UHD Ab IFNg samples differ significantly from samples that contain no UHD Ab IFNg (controls). A linear relationship between the measured parameter and the number of UHD Ab IFNg tablets used was also determined. Therefore, the method meets the validation criterion for linearity.The method has also been shown to provide precision and robustness. Therefore, according to the main criteria (specificity, detection limit, linearity precision and robustness) the method suggested for the identification and quantitative determination of UHD Ab IFNg presence in test samples is valid and can be applied for these purposes. Data processing is performed by means of the R language using the RStudio 1.2.5033 package. The initial fluorescence spectra are smoothed (interpolated) by splines. For this, a cubic spline with 42 nodes is used (the number of nodes is eight times less than the total number of points in the spectrum). The initial data are transformed according to a new basis, which leads to a smoothing of the initial curve. Local peaks are found as points of sign change of the second derivative of the transformed spectrum. Global peaks are selected among those local in proximity to the theoretical positions of the main peaks in the fluorescence spectrum. The data were analyzed statistically using one-way (assessment of specificity) or two-way (assessment of robustness) ANOVA, followed by Tukey’s post-hoc test for pairwise comparisons. The significance level (p) was set at 0.05.本数据集收录了超高稀释效应相关研究的实验结果。当体系中存在抗干扰素γ（Interferon Gamma, IFNg）的超高稀释抗体（Ultrahigh Dilutions of Antibodies, UHD Ab）时，乳糖水溶液的物理化学性质（进而影响其光谱特性）会发生改变。本研究对含有抗干扰素γ的超高稀释抗体的溶液开展了荧光光谱法（Spectrofluorimetry）检测，并测定了其与对照组样品在420nm发射峰强度上的定量差异。该方法的验证研究表明，所提出的检测方法具有特异性：含有抗干扰素γ的超高稀释抗体的样品与不含该类抗体的对照组样品之间存在显著差异。此外，研究还确定了待测参数与所用抗干扰素γ的超高稀释抗体片剂数量之间的线性关系，因此该方法符合线性性验证标准。该方法同时被证实具备精密度与稳健性。综上，依据特异性、检测限、线性性、精密度及稳健性等核心验证准则，本研究提出的用于鉴定并定量测定待测样品中抗干扰素γ的超高稀释抗体含量的方法是可靠有效的，可应用于上述研究目的。本研究采用R语言及RStudio 1.2.5033软件完成数据处理。原始荧光光谱通过样条插值进行平滑处理：具体使用含42个节点的三次样条（Cubic Spline），其节点数量为光谱总点数的1/8，通过将原始数据转换至新的基函数空间以实现原始曲线的平滑。通过变换后光谱的二阶导数符号变化点识别局部峰，并在荧光光谱主峰理论位置附近的局部峰中筛选得到全局峰。数据统计分析采用单因素方差分析（Analysis of Variance, ANOVA）或双因素方差分析，随后通过Tukey事后检验（Tukey’s Post-hoc Test）进行组间两两比较，显著性水平（p值）设定为0.05。