Aljnabi Aws, Hagadorn Kelly A, Neumann Louisa E, Peterson Mary E, Scott Bethany, Miura Kazutoyo, Ongoiba Aissata, Doumbo Safiatou, Doumtabe Didier, Li Shanping, Kayentao Kassoum, Traore Boubacar, Bolland Silvia, Crompton Peter D, Hopp Christine S
B Cell Immunology in Malaria, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
Malaria Infection Biology and Immunity Section, Laboratory of Immunogenetics, Division of Intramural Research, NIAID, NIH, Rockville, MD, USA; Yale School of Public Health, Department of Epidemiology of Microbial Diseases, New Haven, CT, USA.
STAR Protoc. 2025 Jun 20;6(2):103730. doi: 10.1016/j.xpro.2025.103730. Epub 2025 Apr 8.
Autoantibodies (AAbs) contribute to various immune-mediated diseases and are valuable biomarkers for diagnosis, classification, and disease activity. Here, we present a protocol for the affinity enrichment of AAbs from human plasma samples. We describe steps to generate a human cell line lysate, which is immobilized on Sepharose beads for affinity enrichment of AAbs. We then detail the quality-control procedure of verifying autoreactivity of AAb fractions. This protocol has potential application in functional and proteomic analyses of AAbs. For complete details on the use and execution of this protocol, please refer to Hagadorn et al..
自身抗体（AAbs）参与多种免疫介导的疾病，是用于诊断、分类和疾病活动的有价值的生物标志物。在此，我们展示了一种从人血浆样本中亲和富集自身抗体的方案。我们描述了生成人细胞系裂解物的步骤，该裂解物固定在琼脂糖珠上用于自身抗体的亲和富集。然后我们详细说明了验证自身抗体组分自身反应性的质量控制程序。该方案在自身抗体的功能和蛋白质组学分析中具有潜在应用。有关该方案使用和执行的完整详细信息，请参考哈加多恩等人的研究。
