Al-Hawat Marie-Lynn, Caron Justine, Djebbar Sarah, Matoori Simon
Faculté de Pharmacie, Université de Montréal, Montreal, Quebec H3T 1J4, Canada.
ACS Bio Med Chem Au. 2024 Jul 30;4(5):226-232. doi: 10.1021/acsbiomedchemau.4c00013. eCollection 2024 Oct 16.
Ammonia is a key biomarker in inborn and acquired liver disease. As clinical point-of-care blood ammonia assays are lacking, we developed a polymersome formulation for point-of-care blood ammonia sensing combined with a portable fluorometer. A pH-sensitive near-infrared (NIR) fluorescent dye was identified, which showed a strong fluorescence increase at acidic pH values. Building on reports on ammonia-selective PS--PEG polymersomes, these polymersomes were loaded with the NIR dye. These NIR fluorescent polymersomes sensed ammonia in a clinically relevant range in ammonia-spiked fresh whole blood with high linearity ( = 0.9948) after 5 min using a conventional tabletop plate reader. Subsequently, the assay was tested with a portable fluorometer. An ammonia-dependent fluorescence increase was detected in ammonia-spiked fresh mouse blood after 5 min using the portable fluorometer. The NIR dye-loaded PS--PEG polymersomes rapidly sensed ammonia with high linearity in whole blood. This assay was successfully combined with a portable fluorometer and only required 3 μL of blood. These findings motivate a further development and clinical translation of this point-of-care blood ammonia assay.
氨是先天性和后天性肝病的关键生物标志物。由于缺乏临床即时检测血氨的方法，我们开发了一种用于即时检测血氨传感的聚合物囊泡制剂，并结合了便携式荧光计。我们鉴定了一种对pH敏感的近红外（NIR）荧光染料，该染料在酸性pH值下荧光强度显著增加。基于关于氨选择性聚（苯乙烯-共-乙二醇）（PS-PEG）聚合物囊泡的报道，这些聚合物囊泡被装载了近红外染料。使用传统台式酶标仪，这些近红外荧光聚合物囊泡在加氨的新鲜全血中，5分钟后在临床相关氨浓度范围内以高线性度（R² = 0.9948）检测到氨。随后，使用便携式荧光计对该检测方法进行测试。使用便携式荧光计在加氨的新鲜小鼠血液中5分钟后检测到氨依赖性荧光增加。装载近红外染料的PS-PEG聚合物囊泡在全血中能快速且高线性地检测氨。该检测方法成功地与便携式荧光计相结合，且仅需3微升血液。这些发现推动了这种即时检测血氨方法的进一步开发和临床转化。
