Enabling quantification of protein concentration in human serum biopsies using attenuated total reflectance - Fourier transform infrared (ATR-FTIR) spectroscopy
Enabling quantification of protein concentration in human serum biopsies using attenuated total reflectance - Fourier transform infrared (ATR-FTIR) spectroscopy
复制标题
DOI：
10.1016/j.vibspec.2018.08.019
复制
发表时间：
2018-11-01
影响因子：
2.5
通讯作者：
Baker, Matthew J.
中科院分区：
文献类型：
--
作者：
Spalding, Katie;Bonnier, Franck;Baker, Matthew J.
人体血液中蛋白质浓度的变化被用作营养状况、水合作用和潜在疾病的指标。它们通常是在定期的临床预约中测量的，目前的分析过程可能导致等待结果的时间很长，并且需要患者回访。衰减全反射-傅里叶变换红外（ATR-FTIR）光谱有能力检测微小的分子差异，定性和定量，在生物流体样品中，不需要大量的样品制备。ATR-FTIR几乎可以立即返回分析测量结果，因此可以被认为是临床上监测血液分子变化的理想技术。为了确定在临床环境中使用ATR-FTIR光谱进行蛋白质定量的适用性，在分析患者临床样本之前，对混合的人血清样本进行了分析，其中加入了不同浓度的人血清白蛋白（HSA）和免疫球蛋白G （IgG）。使用经过验证的偏最小二乘法，加标样品（IgG）的线性关系高达0.998，RMSEV为0.49 +/- 0.05 mg mL(-1)，患者样品的R-2值为0.992，相应的RMSEV为0.66 +/- 0.05 mg mL（-1）。使用两个盲检验模型验证了这一说法，留下一名患者进行交叉验证和k-fold交叉验证，获得最佳线性和RMSEV值分别为0.934和1.99 +/- 0.79 mg mL（-1）。这表明，ATR-FTIR能够快速且简单地定量临床相关复杂基质和浓度（如血清样品）中的蛋白质。从光谱特征中提供定量步骤以及快速疾病分类的能力将有助于振动光谱的临床翻译，以帮助解决当前患者诊断途径面临的问题。
Changes in protein concentrations within human blood are used as an indicator for nutritional state, hydration and underlying illnesses. They are often measured at regular clinical appointments and the current analytical process can result in long waiting times for results and the need for return patient visits. Attenuated total reflectance- Fourier transform infrared (ATR-FTIR) spectroscopy has the ability to detect minor molecular differences, qualitatively and quantitatively, in biofluid samples, without extensive sample preparation. ATR-FTIR can return an analytical measurement almost instantaneously and therefore could be deemed as an ideal technique for monitoring molecular alterations in blood within the clinic.To determine the suitability of using ATR-FTIR spectroscopy to enable protein quantification in a clinical setting, pooled human serum samples spiked with varying concentrations of human serum albumin (HSA) and immunoglobulin G (IgG) were analysed, before analysing patient clinical samples. Using a validated partial least squares method, the spiked samples (IgG) produced a linearity as high as 0.998 and a RMSEV of 0.49 +/- 0.05 mg mL(-1), with the patient samples producing R-2 values of 0.992 and a corresponding RMSEV of 0.66 +/- 0.05 mg mL(-1). This claim was validated using two blind testing models, leave one patient out cross validation and k-fold cross validation, achieving optimum linearity and RMSEV values of 0.934 and 1.99 +/- 0.79 mg mL(-1), respectively.This demonstrates that ATR-FTIR is able to quantify protein within clinically relevant complex matrices and concentrations, such as serum samples, rapidly and with simple sample preparation. The ability to provide a quantification step, along with rapid disease classification, from a spectroscopic signature will aid clinical translation of vibrational spectroscopy to assist with problems currently faced with patient diagnostic pathways.