Aibara Nozomi, Ohyama Kaname
Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Methods Mol Biol. 2019;2024:333-338. doi: 10.1007/978-1-4939-9597-4_22.
Comprehensive identification and profiling of antigens in immune complexes (IC-antigens) is useful to provide insights into pathophysiology and could form the basis for novel diagnostic and treatment strategies for many immune-related diseases. Immune complexome analysis is the method for comprehensively identifying and profiling IC-antigens in biological fluids (such as serum and cerebrospinal fluid). Here, we describe an IC-antigen detection method; specifically, ICs in biological fluids are captured by using protein G- or protein A-coated beads, are subjected to papain-digestion, elution, and tryptic digestion, and are analyzed by nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS).
全面鉴定和分析免疫复合物中的抗原(IC抗原)有助于深入了解病理生理学,并可为许多免疫相关疾病的新型诊断和治疗策略奠定基础。免疫复合物组分析是一种用于全面鉴定和分析生物体液(如血清和脑脊液)中IC抗原的方法。在此,我们描述一种IC抗原检测方法;具体而言,生物体液中的免疫复合物通过使用蛋白G或蛋白A包被的磁珠捕获,经过木瓜蛋白酶消化、洗脱和胰蛋白酶消化,然后通过纳升液相色谱-串联质谱(nano-LC-MS/MS)进行分析。