Multiplexed biosensor for personal wildfire exposure assessment

总结/摘要： 由于气候变化，野火变得越来越频繁和严重，导致越来越多的人暴露在 主要人口受到生物质燃烧排放的高浓度苯和多环芳烃的影响。 野火暴露对人类的健康和福祉构成严重威胁。目前，评估 个人暴露于野火取决于采集环境样本并使用 昂贵的分析技术，由于时间和环境因素， 代谢物浓度的空间变化。生物标志物的生物检测具有很大的潜力， 克服传统技术的局限性并提供个人暴露的准确测量 程度.在这个SBIR项目中，DL ADV-Tech建议开发一种先进的铕纳米颗粒 增强的时间分辨荧光（TRF）免疫测定生物传感器，其提供极高的灵敏度 (pg/ml水平），周转时间短（<10分钟），操作简单，成本低（每次测试<5美元）， 非侵入性地分析尿液样本中的野火暴露。提出这一建议的一个重大意义 方法是现场和非常准确地分析少量的生物标志物的能力，通过利用我们的 先进的纳米材料信号放大技术和多重传感技术。拟议 生物传感器包括纸基条带，以同时测量多种代谢物，2-萘酚（2-NAP）、1-萘酚（1-NH3）、2-萘酚（2-NH3）、1-NH3。 羟基芘葡糖苷酸（1-OHPG）和S-苯基巯基尿酸（S-PMA）， 为了实现高灵敏度，引入了基于纳米材料铕（Eu）的纳米颗粒（YVO 4：Eu）。便携式 将配备TRF读取器以定量靶标浓度。我们已经证明，使用 先进的Eu基纳米颗粒作为信号标记，所提出的荧光免疫分析预计， 提高尿液中野火相关生物标志物检测的稳定性、可靠性和准确性。并入 通过这种创新的信号放大策略，所提出的生物传感器的灵敏度预计将 显著增加。使用所提出的生物传感器，测量个人野火烟雾暴露 生物标志物可在10分钟内完成，检测限可低至0.1 ng/ml。这项I期 在概念验证项目中，我们将专注于具有两个测试区的生物传感器，同时量化三个 尿代谢产物（2-NAP、1-OHPG和SPMA）。将制作和验证拟议的生物传感器， 将建立生物传感器的分析“品质因数”。在第二阶段，生物传感器将进一步 优化用于真实的世界条件中的现场应用，并扩大用于检测更多的生物标志物 与野火暴露有关。这种高度精确和真正便携的TRF生物传感器将提供一个非- 侵入性、敏感、简单、快速、稳健和廉价评估工具，这将彻底改变当前 生物监测技术，以更短的时间和更低的成本评估个人暴露水平。 1

Summary/Abstract: Wildfire are becoming more frequently and severe due to climate change, resulting in growing exposures of major population to high level of benzene and polycyclic aromatic hydrocarbons from biomass-burning emissions. Wildfire exposure pose a serious threat to the health and the well-being of humans. Currently, assessment of personal exposure to wildfire is dependent on taking environmental samples and analyzing the samples using expensive analytical techniques, which cannot adequately represent personal exposure due to the temporal and spatial variation of the metabolite concentrations. Biological detection of biomarkers has great potential to overcome the limitation of conventional techniques and provides accurate measurements of personal exposure levels. In this SBIR project, DL ADV-Tech proposes to develop an advanced europium-nanoparticles enhanced time-resolved fluorescent (TRF) immunoassay biosensor which provides extremely sensitivity (pg/ml level), short turnaround time (<10 minutes), simple operation and low cost (<$5 for each test) for non-invasively analyzing wildfire exposure in urine samples. A major significance of this proposed approach is the ability to on-site and very accurately analyze low amount of biomarkers by leveraging our advanced nanomaterials signal amplification technique and multiplex sensing technique. The proposed biosensor includes a paper-based strip to simultaneously measure multiple metabolites, 2-naphthol (2-NAP), 1- hydroxypyrene glucuronide (1-OHPG) and S-phenyl mercapturic acid (S-PMA), where newly developed nanomaterial, europium (Eu)-based nanoparticles (YVO4:Eu) is introduced to achieve high sensitivity. A portable TRF reader will be equipped to quantify the concentrations of targets. We have demonstrated that using the advanced Eu-based nanoparticles as signal label, the proposed fluorescent immunoassay is expected to improve the stability, reliability and accuracy in detection of wildfire-associated biomarkers in urine. Incorporated with this innovative signal amplification strategy, the sensitivity of the proposed biosensor is expected to be significantly increased. Using the proposed biosensor, the measurement of personal wildfire smoke exposure biomarkers can be completed within 10 minutes and the detection limit can as low as 0.1 ng/ml. In this Phase I project of proof-of-concept, we will focus on the biosensor with two test zones that simultaneously quantify three urinary metabolites (2-NAP, 1-OHPG and SPMA). The proposed biosensor will be fabricated and validated, and the analytical 'figures of merits' of the biosensor will be established. In Phase II, the biosensor will be further optimized for on-site applications in real world conditions and scaled-up for detecting more biomarkers associated with wildfire exposure. This highly precise and truly portable TRF biosensor will provide a non- invasive, sensitive, simple, rapid, robust, and inexpensive assessment tool, which will revolutionize current biomonitoring technique for assessing personal exposure levels with shorter time and lower cost. 1