Ultra-filtration of human serum for improved quantitative analysis of low molecular weight biomarkers using ATR-IR spectroscopy
Ultra-filtration of human serum for improved quantitative analysis of low molecular weight biomarkers using ATR-IR spectroscopy
复制标题
DOI：
10.1039/c6an01888b
复制
发表时间：
2017-04-21
期刊：
影响因子：
4.2
通讯作者：
Chourpa, Igor
中科院分区：
文献类型：
--
作者：
Bonnier, Franck;Blasco, Helene;Chourpa, Igor
红外光谱是一种可靠、快速、经济的表征技术，可提供样品的分子指纹。预期其灵敏度将能够检测与疾病有关的生物样品中的微小化学变化。ATR-IR特别适用于液体样品分析，尽管通常在数据收集前进行风干，但只需一滴人血清就足以进行筛查和早期诊断。然而，血清成分中生化浓度的动态范围仍然是该技术可靠性的限制因素。利用葡萄糖作为人血清中的峰值模型，本研究表明，在光谱分析之前对血清进行分馏可以显著提高基于偏最小二乘回归算法的定量模型的精度和准确性。通过消耗丰富的高分子量蛋白质（否则会主导收集的光谱特征），监测低分子量成分浓度变化的能力得到增强。验证集（RMSEV）的均方根误差在人血清处理后得到了5倍的改善，预测值的平均相对误差低于1%。此外，该方法很容易转移到不同的体液，这将支持开发更有效和合适的临床方案，用于探索基于振动光谱的离体诊断工具。
Infrared spectroscopy is a reliable, rapid and cost effective characterisation technique, delivering a molecular finger print of the sample. It is expected that its sensitivity would enable detection of small chemical variations in biological samples associated with disease. ATR-IR is particularly suitable for liquid sample analysis and, although air drying is commonly performed before data collection, just a drop of human serum is enough for screening and early diagnosis. However, the dynamic range of constituent biochemical concentrations in the serum composition remains a limiting factor to the reliability of the technique. Using glucose as a model spike in human serum, it has been demonstrated in the present study that fractionating the serum prior to spectroscopic analysis can considerably improve the precision and accuracy of quantitative models based on the partial least squares regression algorithm. By depleting the abundant high molecular weight proteins, which otherwise dominate the spectral signatures collected, the ability to monitor changes in the concentrations of the low molecular weight constituents is enhanced. The Root Mean Square Error for the Validation set (RMSEV) has been improved by a factor of 5 following human serum processing with an average relative error in the predictive values below 1% being achieved. Moreover, the approach is easily transferable to different bodily fluids, which would support the development of more efficient and suitable clinical protocols for exploration of vibrational spectroscopy based ex vivo diagnostic tools.