血浆的自体荧光已被广泛认为是一种前瞻性的疾病筛查方法。然而,对这种内在荧光的评估主要是现象学的,它的起源仍然没有被完全理解,使其在临床实践中的使用复杂化。在这里,我们将详细评估分析能力,可变性,和血浆蛋白荧光的形成基于激发-发射矩阵的开放数据集测量的300名疑似结直肠癌患者,和我们的支持模型实验。使用高分辨率尺寸排阻色谱结合综合光谱分析,我们展示,第一次,HSA在可见光谱范围内(激发波长>350nm)的血浆荧光形成中的主导作用,可能是由其氧化修饰引起的。此外,色氨酸发射的诊断价值,以及蛋白质的酪氨酸荧光和可见荧光通过建立基于树的分类模型来显示,该模型使用一小部分物理可解释的荧光特征来区分对照组和癌症患者,准确率>80%。所获得的结果扩展了当前用于分析血浆荧光的理解和方法,并为新型的基于自发荧光的疾病筛查方法铺平了道路。
Autofluorescence of blood plasma has been broadly considered as a prospective disease screening method. However, the assessment of such intrinsic fluorescence is mostly phenomenological, and its origin is still not fully understood, complicating its use in the clinical practice. Here we present the detailed evaluation of analytical capabilities, variability, and formation of blood plasma protein fluorescence based on the open dataset of excitation-emission matrices measured for ∼300 patients with suspected colorectal cancer, and our supporting model experiments. Using high-resolution size-exclusion chromatography coupled with comprehensive spectral analysis, we demonstrate, for the first time, the dominant role of HSA in the formation of blood plasma fluorescence in the visible spectral range (excitation wavelength >350 nm), presumably caused by its oxidative modifications. Furthermore, the diagnostic value of the tryptophan emission, as well as of the tyrosine fluorescence and visible fluorescence of proteins is shown by building a tree-based classification model that uses a small subset of physically interpretable fluorescence features for distinguishing between the control group and cancer patients with >80% accuracy. The obtained results extend current understanding and approaches used for the analysis of blood plasma fluorescence and pave the way for novel autofluorescence-based disease screening methods.