PDF(Pubmed)
Abstract：
Increased levels of extracellular nicotinamide phosphoribosyltransferase (eNAMPT) are increasingly recognized as a highly useful biomarker of inflammatory disease and disease severity. In preclinical animal studies, a monoclonal antibody that neutralizes eNAMPT has been generated to successfully reduce the extent of inflammatory cascade activation. Thus, the rapid detection of eNAMPT concentration in plasma samples at the point of care (POC) would be of great utility in assessing the benefit of administering an anti-eNAMPT therapeutic. To determine the feasibility of this POC test, we conducted a particle immunoagglutination assay on a paper microfluidic platform and quantified its extent with a flow rate measurement in less than 1 min. A smartphone and cloud-based Google Colab were used to analyze the flow rates automatically. A horizontal flow model and an immunoagglutination binding model were evaluated to optimize the detection time, sample dilution, and particle concentration. This assay successfully detected eNAMPT in both human whole blood and plasma samples (diluted to 10 and 1%), with the limit of detection of 1-20 pg/mL (equivalent to 0.1-0.2 ng/mL in undiluted blood and plasma) and a linear range of 5-40 pg/mL. Furthermore, the smartphone POC assay distinguished clinical samples with low, mid, and high eNAMPT concentrations. Together, these results indicate this POC assay, which utilizes low-cost materials, time-effective methods, and a straightforward immunoassay (without surface immobilization), may reliably allow rapid determination of eNAMPT blood/plasma levels to advantage patient stratification in clinical trials and guide ALT-100 mAb therapeutic decision-making.
摘要：
胞外烟酰胺磷酸核糖基转移酶(eNAMPT)水平的增加越来越被认为是炎性疾病和疾病严重程度的非常有用的生物标志物。在临床前动物研究中，已经产生了中和eNAMPT的单克隆抗体，以成功降低炎症级联激活的程度。因此，在护理点(POC)快速检测血浆样品中的eNAMPT浓度对于评估施用抗eNAMPT治疗剂的益处非常有用.为了确定此POC测试的可行性，我们在纸微流体平台上进行了颗粒免疫凝集测定，并在不到1分钟的时间内通过流速测量对其程度进行了量化。使用智能手机和基于云的GoogleColab自动分析流量。水平流动模型和免疫凝集结合模型进行了评估，以优化检测时间，样品稀释，和颗粒浓度。该测定法成功地检测了人全血和血浆样品中的eNAMPT（稀释至10％和1％），检出限为1-20pg/mL（相当于未稀释血液和血浆中的0.1-0.2ng/mL），线性范围为5-40pg/mL。此外，智能手机POC检测区分临床样本低，mid,和高的eNAMPT浓度。一起,这些结果表明这种POC测定，利用低成本材料，时间有效的方法，和简单的免疫测定（没有表面固定），可以可靠地快速确定eNAMPT血液/血浆水平，以有利于临床试验中的患者分层并指导ALT-100mAb治疗决策。
