Protocol for affinity enrichment of polyclonal autoantibodies from human plasma

Protocol for affinity enrichment of polyclonal autoantibodies from human plasma

Summary: 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 ly...

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Bibliographic Details
Main Authors: Aws Aljnabi, Kelly A. Hagadorn, Louisa E. Neumann, Mary E. Peterson, Bethany Scott, Kazutoyo Miura, Aissata Ongoiba, Safiatou Doumbo, Didier Doumtabe, Shanping Li, Kassoum Kayentao, Boubacar Traore, Silvia Bolland, Peter D. Crompton, Christine S. Hopp
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:STAR Protocols
Subjects:
Cell-based Assays
Immunology
Antibody
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725001364
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Summary:Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667

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