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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| Format: | Article |
| Language: | English |
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Elsevier
2025-06-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725001364 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Aws Aljnabi |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-fedc002f25d34dbd8aabc71abbf6e2ee |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-fedc002f25d34dbd8aabc71abbf6e2ee2025-08-20T02:08:27ZengElsevierSTAR Protocols2666-16672025-06-016210373010.1016/j.xpro.2025.103730Protocol for affinity enrichment of polyclonal autoantibodies from human plasmaAws Aljnabi0Kelly A. Hagadorn1Louisa E. Neumann2Mary E. Peterson3Bethany Scott4Kazutoyo Miura5Aissata Ongoiba6Safiatou Doumbo7Didier Doumtabe8Shanping Li9Kassoum Kayentao10Boubacar Traore11Silvia Bolland12Peter D. Crompton13Christine S. Hopp14B Cell Immunology in Malaria, Bernhard Nocht Institute for Tropical Medicine, Hamburg, GermanyMalaria 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, USAB Cell Immunology in Malaria, Bernhard Nocht Institute for Tropical Medicine, Hamburg, GermanyMalaria Infection Biology and Immunity Section, Laboratory of Immunogenetics, Division of Intramural Research, NIAID, NIH, Rockville, MD, USAAutoimmunity and Functional Genomics Section, Laboratory of Immunogenetics, Division of Intramural Research, NIAID, NIH, Rockville, MD, USALaboratory of Malaria and Vector Research, Division of Intramural Research, NIAID, NIH, Rockville, MD, USAMalaria Research and Training Centre, Department of Epidemiology of Parasitic Diseases, International Center of Excellence in Research, University of Sciences, Technique and Technology of Bamako, Bamako, MaliMalaria Research and Training Centre, Department of Epidemiology of Parasitic Diseases, International Center of Excellence in Research, University of Sciences, Technique and Technology of Bamako, Bamako, MaliMalaria Research and Training Centre, Department of Epidemiology of Parasitic Diseases, International Center of Excellence in Research, University of Sciences, Technique and Technology of Bamako, Bamako, MaliMalaria Infection Biology and Immunity Section, Laboratory of Immunogenetics, Division of Intramural Research, NIAID, NIH, Rockville, MD, USAMalaria Research and Training Centre, Department of Epidemiology of Parasitic Diseases, International Center of Excellence in Research, University of Sciences, Technique and Technology of Bamako, Bamako, MaliMalaria Research and Training Centre, Department of Epidemiology of Parasitic Diseases, International Center of Excellence in Research, University of Sciences, Technique and Technology of Bamako, Bamako, MaliAutoimmunity and Functional Genomics Section, Laboratory of Immunogenetics, Division of Intramural Research, NIAID, NIH, Rockville, MD, USAMalaria Infection Biology and Immunity Section, Laboratory of Immunogenetics, Division of Intramural Research, NIAID, NIH, Rockville, MD, USAB Cell Immunology in Malaria, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166725001364Cell-based AssaysImmunologyAntibody |
| spellingShingle | 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 Protocol for affinity enrichment of polyclonal autoantibodies from human plasma STAR Protocols Cell-based Assays Immunology Antibody |
| title | Protocol for affinity enrichment of polyclonal autoantibodies from human plasma |
| title_full | Protocol for affinity enrichment of polyclonal autoantibodies from human plasma |
| title_fullStr | Protocol for affinity enrichment of polyclonal autoantibodies from human plasma |
| title_full_unstemmed | Protocol for affinity enrichment of polyclonal autoantibodies from human plasma |
| title_short | Protocol for affinity enrichment of polyclonal autoantibodies from human plasma |
| title_sort | protocol for affinity enrichment of polyclonal autoantibodies from human plasma |
| topic | Cell-based Assays Immunology Antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725001364 |
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