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We report the discovery and development of a synthetic human Fab as the first immunoPET tracer targeting human CD68 (huCD68), the gold standard histopathological biomarker of pan-macrophages.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Phage display was used to identify novel huCD68-specific Fabs. The lead Fab for immunoPET development was chosen based on expression yield and stability. Fab was conjugated to p-SCN-Bn-DFO and radiolabeled with\n                      <jats:sup>89<\/jats:sup>\n                      Zr, yielding [\n                      <jats:sup>89<\/jats:sup>\n                      Zr]Zr-DFO-Fab. Stability and specific binding of the tracer to THP-1 macrophages were assessed in vitro. The specificity of the tracer in vivo was characterized by PET imaging of two different animal models: 1) BALB\/c mice bearing huCD68 antigen- or irrelevant protein-plugs, and 2) tumor xenografted NSG mice intratumorally injected with either THP-1 macrophages or THP-1 undifferentiated cells. Finally, tumors were subjected to immunofluorescence (IF) staining of huCD68 and correlated with PET standardized uptake values (SUV).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Four novel huCD68-specific Fabs were identified. Fab8 was chosen as the lead based on high expression yield, purity, and stability. In vivo, [\n                      <jats:sup>89<\/jats:sup>\n                      Zr]Zr-DFO-Fab8 uptake was significantly higher in huCD68 antigen-plugs than in irrelevant protein-plugs at 1- and 4-h post-injection (p.i.). Tracer uptake in THP-1 macrophage tumors was also significantly higher than in THP-1 undifferentiated tumors at 1- and 6-h p.i.. IF of tumors confirmed higher density of huCD68 in those implanted with THP-1 macrophages than those with THP-1 undifferentiated cells. CD68 expression highly correlated with immunoPET SUV.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      [\n                      <jats:sup>89<\/jats:sup>\n                      Zr]Zr-DFO-Fab8 binds specifically to huCD68, supporting the feasibility of direct PET-to-histopathogical correlation of total TAM burden.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s00259-026-08180-y","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T06:50:33Z","timestamp":1790578233000},"page":"7770-7785","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A FABulous approach to image human macrophages"],"prefix":"10.1007","volume":"53","author":[{"given":"Borna","family":"Roohani","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7055-0413","authenticated-orcid":false,"given":"Ayla","family":"Vaughn Embs","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin","family":"Kumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Supum","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha R.","family":"Katz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chloe","family":"La Prairie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dajah","family":"Nash","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mann","family":"Dangarwala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harriet M.","family":"Kluger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daryl E.","family":"Klein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin M.","family":"Larimer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bryce","family":"Nelson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9024-0750","authenticated-orcid":false,"given":"Bernadette","family":"Marquez-Nostra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,28]]},"reference":[{"key":"8180_CR1","doi-asserted-by":"publisher","DOI":"10.1155\/2017\/9624760","author":"SR Nielsen","year":"2017","unstructured":"Nielsen SR, Schmid MC. 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Therefore, informed consent to participate was not required.","order":2,"name":"Ethics","label":"Consent to participate","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"No individual person\u2019s data, images, or identifiable information are included in this manuscript. Therefore, consent to publish was not required.","order":3,"name":"Ethics","label":"Consent to publish","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Author BMN has received research reagents from Janssen Pharmaceuticals at no charge. Author HMK has received research support from the following: 1) Institutional Research Grants- Merck, Bristol-Myers Squibb, Apexigen, Pfizer, Asherbio Therapeutics 2) Personal Fees- Iovance, Merck, Chemocentryx, Bristol-Myers Squibb, Gigagen, GI reviewers, Pliant Therapeutics, Esai, Invox, Wherewolf, Teva, Replimmune, Genmab, Immunocore, TuHura. Author BN has received funding from Gilead Sciences and is currently an employee of Orion Pharma. BML is an employee of and holds equity in Cytosite Bio.","order":4,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}}]}}