Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Campus de Cantoblanco, Madrid, Spain.
Talanta. 2013 May 15;109:209-15. doi: 10.1016/j.talanta.2013.02.018. Epub 2013 Feb 13.
Surface plasmon resonance (SPR)-based biosensors are established tools for measuring biomolecular interactions between unlabeled analytes in real time, and are thus an ideal method to evaluate G protein-coupled receptor (GPCR) binding interactions. Using as a vehicle lentiviral particles bearing the chemokine receptor CXCR4 in its native plasma membrane context, SPR analysis can be performed using the particles as specific receptors to monitor the CXCR4 interaction with its ligand, CXCL12. The method shows linear correlation in the 5-40 nM range, with low intra- and inter-assay variation, a relative standard deviation <10%, chip-to-chip variation <12%, with stability of the sensor response for more than 150 measurements in the same chip over a four-week period. Our objective was to develop a method for rapid detection and quantification of analytes such as CXCL12 in biological samples, with no need for pretreatment. As a proof of concept, we tested for CXCL12 in urine samples from rheumatoid arthritis patients, who have elevated levels of this chemokine in plasma and synovial fluid. The biosensor method allowed sensitive, reproducible CXCL12 detection in the physiological range, suggesting its value for the diagnosis of autoimmune disorders.
基于表面等离子体共振(SPR)的生物传感器是实时测量未标记分析物之间生物分子相互作用的成熟工具,因此是评估 G 蛋白偶联受体(GPCR)结合相互作用的理想方法。使用带有天然质膜环境中趋化因子受体 CXCR4 的慢病毒颗粒作为载体,SPR 分析可以使用颗粒作为特异性受体来监测 CXCR4 与配体 CXCL12 的相互作用。该方法在 5-40 nM 范围内呈线性相关,具有低的内和间分析变异,相对标准偏差 <10%,芯片间变异 <12%,在同一张芯片上进行超过 150 次测量的传感器响应稳定性超过 4 周。我们的目标是开发一种快速检测和定量生物样品中分析物(如 CXCL12)的方法,无需预处理。作为概念验证,我们测试了类风湿关节炎患者尿液样本中的 CXCL12,这些患者的这种趋化因子在血浆和滑液中含量升高。生物传感器方法允许在生理范围内灵敏、可重复地检测 CXCL12,表明其对自身免疫性疾病诊断的价值。