{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T12:54:22Z","timestamp":1764939262498,"version":"3.46.0"},"reference-count":40,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2025,11,6]],"date-time":"2025-11-06T00:00:00Z","timestamp":1762387200000},"content-version":"vor","delay-in-days":5,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2025,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Unattended endotoxin (ETX) contamination in biological samples constitute a major challenge for\n                    <jats:italic>in vitro<\/jats:italic>\n                    and\n                    <jats:italic>in vivo<\/jats:italic>\n                    applications. Besides being potentially life\u2010threatening, ETX contamination is especially relevant in global transcriptome analyses, where competing ETX stimulation can significantly skew the final gene expression profile. Our studies in mice and cultured skin epithelial cells (epidermal keratinocytes) aiming to characterize the effect of antibodies such as AK23 immunoglobulins (IgG directed against the cell\u2010cell adhesion molecule desmoglein [DSG] 3) in the autoimmune disease pemphigus vulgaris (PV) revealed that laboratory\u2010produced and even commercial control antibodies can exhibit non\u2010negligible ETX contaminations. Moreover, these contaminants are extremely difficult to remove. To overcome these challenges, we have devised a simple yet nontoxic and scalable two\u2010step protocol to efficiently reduce ETX levels during or after the IgG purification process. It consists firstly of 0.5 M NaOH pre\u2010treatment of all devices, including the protein A resin, used during IgG sanitization and purification, in parallel with meticulous in\u2010process monitoring of ETX levels. Secondly, before IgG elution from protein A, ETX is stripped from IgG by ion\u2010exchange with the common amino acid arginine. This two\u2010step approach successfully reduces ETX by &gt;95% from hybridoma\u2010derived, laboratory\u2010produced AK23 IgG, as well as patient PV and control IgG, resulting in an 85% IgG recovery rate and ETX levels compatible with\n                    <jats:italic>U.S. Pharmacopeia<\/jats:italic>\n                    guidelines. \u00a9 2025 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Sanitization of devices and protein A resin with\n                    <jats:sc>0.5 M NaOH<\/jats:sc>\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : On\u2010column stripping of ETX from AK23 IgG\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Quality control of sanitized AK23 IgG\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70238","type":"journal-article","created":{"date-parts":[[2025,11,6]],"date-time":"2025-11-06T12:20:06Z","timestamp":1762431606000},"source":"Crossref","is-referenced-by-count":0,"title":["Efficient On\u2010Column Removal of Endotoxin from Immunoglobulins Such as AK23"],"prefix":"10.1002","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7631-8005","authenticated-orcid":false,"given":"Siavash","family":"Rahimi","sequence":"first","affiliation":[{"name":"Department for Biomedical Research, Molecular Dermatology and Stem Cell Research University of Bern  Bern Switzerland"},{"name":"Department of Dermatology, Inselspital, Bern University Hospital University of Bern  Bern Switzerland"},{"name":"Graduate School Cellular and Biomedical Sciences University of Bern  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-0129-4504","authenticated-orcid":false,"given":"Patrizia","family":"Sauta","sequence":"additional","affiliation":[{"name":"Department for Biomedical Research, Molecular Dermatology and Stem Cell Research University of Bern  Bern Switzerland"},{"name":"Department of Dermatology, Inselspital, Bern University Hospital University of Bern  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5498-1142","authenticated-orcid":false,"given":"Monika","family":"Edler","sequence":"additional","affiliation":[{"name":"CSL, CSL Biologics Research Centre  Bern Switzerland"},{"name":"Swiss Institute for Translational and Entrepreneurial Medicine, sitem\u2010insel  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3521-4459","authenticated-orcid":false,"given":"Elisabeth","family":"Locher","sequence":"additional","affiliation":[{"name":"CSL, CSL Biologics Research Centre  Bern Switzerland"},{"name":"Swiss Institute for Translational and Entrepreneurial Medicine, sitem\u2010insel  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5984-1819","authenticated-orcid":false,"given":"Marlies","family":"Illi","sequence":"additional","affiliation":[{"name":"CSL, CSL Biologics Research Centre  Bern Switzerland"},{"name":"Swiss Institute for Translational and Entrepreneurial Medicine, sitem\u2010insel  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6694-7762","authenticated-orcid":false,"given":"Taravat","family":"Shojaeian","sequence":"additional","affiliation":[{"name":"Department for Biomedical Research, Molecular Dermatology and Stem Cell Research University of Bern  Bern Switzerland"},{"name":"Department of Dermatology, Inselspital, Bern University Hospital University of Bern  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0424-6297","authenticated-orcid":false,"given":"Luca","family":"Borradori","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Inselspital, Bern University Hospital University of Bern  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-9470-9877","authenticated-orcid":false,"given":"Thomas","family":"Gentinetta","sequence":"additional","affiliation":[{"name":"CSL, CSL Biologics Research Centre  Bern Switzerland"},{"name":"Swiss Institute for Translational and Entrepreneurial Medicine, sitem\u2010insel  Bern Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4749-1858","authenticated-orcid":false,"given":"William V. 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