Real-Fernández Feliciana, Rossi Giada, Lolli Francesco, Papini Anna Maria, Rovero Paolo
Interdepartmental Laboratory of Peptide and Protein Chemistry and Biology-PeptLab, Italy ; Department of Neurosciences, Psychology, Drug Research and Child Health, Section of Pharmaceutical Sciences and Nutraceutics, University of Florence, Via Ugo Schiff 6, I-50019 Sesto Fiorentino, Italy.
Interdepartmental Laboratory of Peptide and Protein Chemistry and Biology-PeptLab, Italy ; Department of Biomedical, Experimental and Clinical Sciences, University of Florence, Viale Morgagni 50, I-50134 Firenze, Italy.
MethodsX. 2015 Mar 12;2:141-4. doi: 10.1016/j.mex.2015.03.004. eCollection 2015.
Surface plasmon resonance technique is particularly interesting in immunology because it has the potential to visualize label-free antigen-antibody interactions in real-time, thus enabling antibody detection and monitoring. Herein we release the guidelines for the correct use of a method to detect specific antibodies directly in Multiple Sclerosis patients' sera using a glucopeptide-based label-free biosensor. The protocol describes the strategy employed for the immobilization of glucopeptide antigen onto a gold sensor chip and the evaluation of the specific binding of serum antibodies to the immobilized antigen. •Label-free method for the real time screening of disease-specific antibodies within a few minutes;•The described protocol employs small quantities of glucopeptide antigen and blood serum samples saving method-cost;•Stability of the immobilized glucopeptide antigen guarantees the regeneration of the surface allowing re-use the immunosensor with high automated throughput. The antibodies detected using the described methodology can be evaluated as biomarkers of Multiple Sclerosis. The SPR detection system is able to characterize antibodies significantly different from those evaluated in the classical enzyme-linked immunosorbent assays (ELISA).
表面等离子体共振技术在免疫学中特别有趣，因为它有潜力实时可视化无标记的抗原-抗体相互作用，从而实现抗体检测和监测。在此，我们发布了使用基于糖肽的无标记生物传感器直接检测多发性硬化症患者血清中特异性抗体的正确方法指南。该方案描述了将糖肽抗原固定到金传感器芯片上所采用的策略，以及评估血清抗体与固定抗原的特异性结合。•无标记方法可在几分钟内实时筛选疾病特异性抗体；•所述方案使用少量糖肽抗原和血清样本，节省方法成本；•固定化糖肽抗原的稳定性保证了表面的再生，允许以高自动化通量重复使用免疫传感器。使用所述方法检测到的抗体可作为多发性硬化症的生物标志物进行评估。表面等离子体共振检测系统能够表征与经典酶联免疫吸附测定（ELISA）中评估的抗体显著不同的抗体。
