Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Adv Clin Chem. 2013;60:129-41. doi: 10.1016/b978-0-12-407681-5.00004-0.
Immune complexes (ICs) are produced during an immune response and may reflect some aspects of an ongoing immune response. Therefore, the identity of antigens incorporated into ICs provides the information that in the future may aid in the development of diagnosis and treatment strategies for autoimmune diseases, infection, cancer, and transplantation therapy, and this information might be more relevant than the information on free antigens. Because ICs may contain many antigens, comprehensive identification and profiling of such antigens are more effective than immunoblotting detection. Here, we introduced mass spectrometry (MS)-based two approaches (immunoproteomics and immune complexome analysis) to comprehensively identify the antigens. Immunoproteomics is a concept to identify disease-associated antigens that elicit immune responses by combining protein separation (two-dimensional electrophoresis, gel-free separation), immunological detection (Western blotting), and MS or by combining immunocapture and MS. Immune complexome analysis is designed for identifying antigens in circulating ICs and consists of ICs separation from serum and direct tryptic digestion followed by nano-liquid chromatography-tandem MS.
免疫复合物（ICs）是在免疫反应过程中产生的，可能反映了正在进行的免疫反应的某些方面。因此，包含在 ICs 中的抗原的身份提供了未来可能有助于开发自身免疫性疾病、感染、癌症和移植治疗的诊断和治疗策略的信息，这些信息可能比游离抗原的信息更相关。由于 ICs 可能包含许多抗原，因此全面鉴定和分析这些抗原比免疫印迹检测更有效。在这里，我们介绍了基于质谱（MS）的两种方法（免疫蛋白质组学和免疫复合物组学分析）来全面鉴定抗原。免疫蛋白质组学是一种通过结合蛋白质分离（二维电泳、无胶分离）、免疫检测（Western blot）和 MS 或通过结合免疫捕获和 MS 来鉴定引起免疫反应的疾病相关抗原的概念。免疫复合物组学是为鉴定循环 IC 中的抗原而设计的，包括从血清中分离 IC 和直接胰蛋白酶消化，然后进行纳流液相色谱-串联 MS。
