Ramos Kenia Chávez, Macías María Del Pilar Cañizares
Laboratorio de Métodos de Flujo Continuo, Departamento de Química Analítica, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico.
Laboratorio de Métodos de Flujo Continuo, Departamento de Química Analítica, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico.
Talanta. 2021 Mar 1;224:121801. doi: 10.1016/j.talanta.2020.121801. Epub 2020 Oct 23.
Anti-cyclic citrullinated peptide IgG antibodies (anti-CCP) are produced as an immune response in the presence of post-translational modified peptides known as cyclic citrullinated peptides (CCP). Anti-CCP have been considered as specific biomarkers for the diagnosis of rheumatoid arthritis (RA), and due to their high specificity, it is possible to make a differential diagnosis of other rheumatic diseases. These autoantibodies can be detected in the early stages of RA and even up to 10 years before presenting the first symptoms of the disease opening a window of opportunity for timely treatment. In this work, a simple straight channel microdevice and CCP conjugated magnetic nanoparticles (MNPs-CCP) as solid support was developed for quantifying anti-CCP. An additional microdevice with an optical flow Z cell design coupled with optical fibers was used to perform the spectrophotometric detection. The dynamic range of concentrations was determined between 0.70 and 2000 U mL with a limit of detection of 0.70 U mL. The developed microdevice immunoassay was probed using a positive control and a negative control of plasma employing only 6 μL of both samples and reagents. The results showed that the proposed microdevice was almost nine times faster than using a commercial anti-CCP ELISA kit obtaining equivalent results and being 16 times more sensitive. The microdevice immunoassay, with conjugated MNPs-CCP is a simple method for anti-CCP quantification being cheaper, faster, and more sensitive than the ELISA kit.
抗环瓜氨酸肽IgG抗体(抗CCP)是在存在称为环瓜氨酸肽(CCP)的翻译后修饰肽的情况下作为免疫反应产生的。抗CCP被认为是类风湿性关节炎(RA)诊断的特异性生物标志物,并且由于其高特异性,有可能对其他风湿性疾病进行鉴别诊断。这些自身抗体可以在RA的早期阶段检测到,甚至在出现该疾病的首个症状前10年就能检测到,这为及时治疗打开了机会之窗。在这项工作中,开发了一种简单的直通道微装置以及作为固体支持物的CCP偶联磁性纳米颗粒(MNPs-CCP)用于定量抗CCP。另一个带有光流Z细胞设计并与光纤耦合的微装置用于进行分光光度检测。确定浓度的动态范围在0.70至2000 U/mL之间,检测限为0.70 U/mL。仅使用6μL的样品和试剂,采用血浆的阳性对照和阴性对照对所开发的微装置免疫测定法进行了检测。结果表明,所提出的微装置比使用商业抗CCP ELISA试剂盒快近9倍,获得了等效结果且灵敏度高16倍。带有偶联MNPs-CCP的微装置免疫测定法是一种用于抗CCP定量的简单方法,比ELISA试剂盒更便宜、更快且更灵敏。