{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T12:16:56Z","timestamp":1792671416317,"version":"4.5.0"},"reference-count":78,"publisher":"Wiley","issue":"19","license":[{"start":{"date-parts":[[2017,9,11]],"date-time":"2017-09-11T00:00:00Z","timestamp":1505088000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["ERC\u20102011\u2010AdG 294709\u2010DAMAGECONTROL"],"award-info":[{"award-number":["ERC\u20102011\u2010AdG 294709\u2010DAMAGECONTROL"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["1003484"],"award-info":[{"award-number":["1003484"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2017,10]]},"abstract":"<jats:p>\n                    Extracellular hemoglobin, a byproduct of hemolysis, can release its prosthetic heme groups upon oxidation. This produces metabolically active heme that is exchangeable between acceptor proteins, macromolecules and low molecular weight ligands, termed here labile heme. As it accumulates in plasma labile heme acts in a pro\u2010oxidant manner and regulates cellular metabolism while exerting pro\u2010inflammatory and cytotoxic effects that foster the pathogenesis of hemolytic diseases. Here, we developed and characterized a panel of heme\u2010specific single domain antibodies (sdAbs) that together with a cellular\u2010based heme reporter assay, allow for quantification and characterization of labile heme in plasma during hemolytic conditions. Using these approaches, we demonstrate that when generated during hemolytic conditions labile heme is bound to plasma molecules with an affinity higher than 10\n                    <jats:sup>\u22127<\/jats:sup>\n                    <jats:sc>m<\/jats:sc>\n                    and that 2\u20138% (~ 2\u20135 \u03bc\n                    <jats:sc>m<\/jats:sc>\n                    ) of the total amount of heme detected in plasma can be internalized by bystander cells, termed here bioavailable heme. Acute, but not chronic, hemolysis is associated with transient reduction of plasma heme\u2010binding capacity, that is, the ability of plasma molecules to bind labile heme with an affinity higher than 10\n                    <jats:sup>\u22127<\/jats:sup>\n                    <jats:sc>m<\/jats:sc>\n                    . The heme\u2010specific sdAbs neutralize the pro\u2010oxidant activity of soluble heme\n                    <jats:italic>in vitro<\/jats:italic>\n                    , suggesting that these maybe used to counter the pathologic effects of labile heme during hemolytic conditions. Finally, we show that heme\u2010specific sdAbs can be used to visualize cellular heme. In conclusion, we describe a panel of heme\u2010specific sdAbs that when used with other approaches provide novel insights to the pathophysiology of heme.\n                  <\/jats:p>","DOI":"10.1111\/febs.14192","type":"journal-article","created":{"date-parts":[[2017,8,7]],"date-time":"2017-08-07T08:46:37Z","timestamp":1502095597000},"page":"3278-3301","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":77,"title":["Characterization of plasma labile heme in hemolytic conditions"],"prefix":"10.1111","volume":"284","author":[{"given":"Z\u00e9lia","family":"Gouveia","sequence":"first","affiliation":[{"name":"Instituto Gulbenkian da Ci\u00eancia Oeiras Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana R.","family":"Carlos","sequence":"additional","affiliation":[{"name":"Instituto Gulbenkian da Ci\u00eancia Oeiras Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojing","family":"Yuan","sequence":"additional","affiliation":[{"name":"Department of Animal and Avian Sciences and Department of Cell Biology and Molecular Genetics University of Maryland College Park MD USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederico","family":"Aires\u2010da\u2010Silva","sequence":"additional","affiliation":[{"name":"Technophage S.A. 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