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Microfluidic device for single step measurement of protein C in plasma samples for sepsis prognosis
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-05-30 , DOI: 10.1039/d1lc01084k
Sreekant Damodara 1 , Jaskirat Arora 2 , Dhruva J Dwivedi 2 , Patricia C Liaw 2 , Alison E Fox-Robichaud 2 , P Ravi Selvaganapathy 1 , Canadian Critical Care Translational Biology Group
Affiliation  

Protein C is a vitamin K dependant protein in plasma that plays an essential role in regulating the coagulation cascade and inflammatory response. As a result of its importance in these roles, it has been suggested as a biomarker for prognosis of patients affected by sepsis. Sepsis is a dysregulated host response to an infection that is the leading cause of mortality in U.S. hospitals and results in the highest cost of hospitalization. It was found that protein C concentration in non-surviving sepsis patients is significantly lower (1.8 μg mL−1) than in survivors and healthy patients who have a protein C concentration of 3.9–5.9 μg mL−1. Current methods for diagnosing sepsis rely on expensive immunoassays or functional assays that require multiple steps for isolation and activation of protein C. We demonstrate in this paper a low cost, single step assay for detection of protein C in blood plasma. This was done by combining isoelectric gates with barium-immobilized metal affinity trapping. The electric field was optimized for use with immobilized metal affinity using COMSOL simulation. The integrated device was tested with samples containing buffered protein C, protein C in the presence of high concentration bovine serum albumin and alpha 1-proteinase inhibitor, and in blood plasma with spiked protein C. The stability of the measured values was tested by monitoring the intensity of a mixture of protein C with BSA and A1PI every minute to determine that measurement after 40 minutes was optimal. The results showed that the device could be used to distinguish a reduction in protein C from 4.46 μg mL−1 to 1.96 μg mL−1 with greater than 98% confidence in plasma making it suitable for sepsis prognosis.

中文翻译:

用于脓毒症预后的血浆样品中蛋白 C 单步测量的微流控装置

蛋白 C 是血浆中的一种维生素 K 依赖性蛋白,在调节凝血级联反应和炎症反应中起重要作用。由于其在这些作用中的重要性,已建议将其作为脓毒症患者预后的生物标志物。脓毒症是宿主对感染的失调反应,感染是美国医院死亡的主要原因,并导致住院费用最高。发现非存活脓毒症患者的蛋白 C 浓度(1.8 μg mL -1 )显着低于蛋白 C 浓度为 3.9-5.9 μg mL -1的幸存者和健康患者. 目前诊断败血症的方法依赖于昂贵的免疫测定或功能测定,需要多个步骤来分离和激活蛋白 C。我们在本文中展示了一种低成本、单步检测血浆中蛋白 C 的方法。这是通过将等电门与钡固定金属亲和捕获相结合来完成的。使用 COMSOL 仿真对电场进行了优化,以用于固定金属亲和性。使用含有缓冲蛋白 C 的样品、在高浓度牛血清白蛋白和 α 1-蛋白酶抑制剂存在下的蛋白 C 以及在加标蛋白 C 的血浆中对集成装置进行了测试。通过每分钟监测蛋白质 C 与 BSA 和 A1PI 的混合物的强度来测试测量值的稳定性,以确定 40 分钟后的测量是最佳的。结果表明,该装置可用于区分蛋白质 C 的减少与 4.46 μg/mL-1至 1.96 μg mL -1对血浆的置信度大于 98%,使其适用于脓毒症预后。
更新日期:2022-05-30
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