Identification of neuronal surface autoantigens in serum samples from a patient with thymoma-associated limbic encephalitis, comparison of pre- and postoperative thymoma by LC-MS/MS analysis

Mouse brain tissue lysates were immunoprecipitated with protein A/G Sepharose beads which had been incubated with serum from patient or control participant. The immune complexes were subsequently separated by SDS‒PAGE. Bands were excised from the gel, and trypsin-digested samples were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) using an Orbitrap Velos Pro (Thermo Scientific). MS/MS dataset analysis was performed using the Mascot software program (Matrix Science).

小鼠脑组织裂解物经与患者或对照受试者血清孵育后的蛋白A/G琼脂糖凝胶珠（protein A/G Sepharose beads）进行免疫沉淀。所得免疫复合物随后经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离。从凝胶中切取电泳条带，经胰蛋白酶消化后的样品采用Orbitrap Velos Pro（赛默飞世尔科技，Thermo Scientific）仪器开展液相色谱-串联质谱（LC-MS/MS）分析。质谱二级图谱数据集的分析工作采用Mascot软件（Matrix Science）完成。