Department of Mechanical and Biomedical Engineering, Ewha Womans University, Seoul, 03760, Republic of Korea; Graduate Program in Smart Factory, Ewha Womans University, Seoul, 03760, Republic of Korea.
Department of Neurology, Seoul Hospital, College of Medicine, Ewha Womans University, Seoul, 07804, Republic of Korea.
Anal Chim Acta. 2024 Dec 1;1331:343347. doi: 10.1016/j.aca.2024.343347. Epub 2024 Oct 16.
As anticoagulants are widely used to treat patients with atrial fibrillation (AF) and other thrombotic conditions, it is necessary for physicians to determine whether the medication has been taken in emergencies. Among many anticoagulants, rivaroxaban has attracted attention due to its safety and convenience. Since rivaroxaban inhibits activated coagulation factor X (factor Xa), measuring factor Xa activity can determine the presence of rivaroxaban.
We report a magnetic biosensing platform capable of measuring the activity of factor Xa using peptide substrates conjugated with magnetic nanoparticles (MNPs). Due to the size of factor Xa, a new method of solution-phase assays was proposed for magnetic biosensing platforms to address issues with immobilized peptides on the sensors. This method was validated with factor Xa and trypsin, both of which are serine proteases. In the solution-phase assays, samples with either the enzymes of interest or no enzyme were simultaneously measured, and the activity of the enzyme was estimated using the difference between the measurements. Unlike conventional optical methods, our platform was capable of detecting the activity of factor Xa at 2 μg mL with a 30 min sample incubation.
The assay using giant magnetoresistive biosensors outperformed conventional optical techniques. This platform can facilitate the determination of the presence of rivaroxaban and assist physicians in deciding on appropriate treatments for patients.
由于抗凝剂被广泛用于治疗心房颤动（AF）和其他血栓性疾病的患者，医生有必要确定药物是否在紧急情况下服用。在许多抗凝剂中，利伐沙班因其安全性和便利性而引起关注。由于利伐沙班抑制激活的凝血因子 X（因子 Xa），因此测量因子 Xa 的活性可以确定利伐沙班的存在。
我们报告了一种使用与磁性纳米颗粒（MNPs）偶联的肽底物来测量因子 Xa 活性的磁生物传感平台。由于因子 Xa 的大小，我们为磁性生物传感平台提出了一种新的溶液相测定方法，以解决传感器上固定化肽的问题。该方法使用因子 Xa 和胰蛋白酶进行了验证，这两种酶都是丝氨酸蛋白酶。在溶液相测定中，同时测量了含有感兴趣酶或无酶的样品，并使用测量值之间的差异来估计酶的活性。与传统的光学方法不同，我们的平台能够在 30 分钟的样品孵育时间内以 2μg/mL 的浓度检测因子 Xa 的活性。
使用巨磁电阻生物传感器的测定方法优于传统的光学技术。该平台可以促进利伐沙班存在的确定，并帮助医生为患者决定适当的治疗方案。
