Protocol for affinity enrichment of polyclonal autoantibodies from human plasma
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.<sup>1</sup>.
Authors
- Mary Peterson (ORCID: https://orcid.org/0000-0001-6848-8709)
- Shanping Li (ORCID: https://orcid.org/0000-0003-0998-2082)
- Didier Doumtabé (ORCID: https://orcid.org/0000-0002-6888-7918)
- Silvia Bolland (ORCID: https://orcid.org/0000-0002-9662-2394)
- Aws Aljnabi
- Kassoum Kayentao (ORCID: https://orcid.org/0000-0001-6877-0093)
- Aïssata Ongoïba (ORCID: https://orcid.org/0000-0002-8797-497X)
- Boubacar Traoré
- Kelly A. Hagadorn (ORCID: https://orcid.org/0000-0001-7200-7023)
- Peter D. Crompton (ORCID: https://orcid.org/0000-0001-5629-0412)
- Safiatou Doumbo (ORCID: https://orcid.org/0000-0002-0974-0860)
- Christine S. Hopp
- Kazutoyo Miura (ORCID: https://orcid.org/0000-0003-4455-2432)
- Bethany Scott (ORCID: https://orcid.org/0000-0003-1009-2458)
- Lily Neumann
Institutions
- University of Bamako (ML)
- Office of Extramural Research (US)
- Institute of Infection and Immunity (CA)
- Bernhard Nocht Institute for Tropical Medicine (DE)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-04-08
- DOI
- https://doi.org/10.1016/j.xpro.2025.103730
- Primary Topic
- Monoclonal and Polyclonal Antibodies Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Division of Intramural Research, National Institute of Allergy and Infectious Diseases
- Deutsche Forschungsgemeinschaft
- National Institutes of Health
- Leibniz-Gemeinschaft
- National Institute of Allergy and Infectious Diseases